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NB100-2220

LC3B Antibody - BSA Free

(51 Reviews)

(1669 Publications)

0.1 ml

NB100-2220

Lysates of HeLa parental cell line and LC3B knockout HeLa cell line (KO) untreated (-) or treated (+) with 50 uM CQ for 18 hours. PVDF (Polyvinylidene difluoride) membrane was probed with 0.5 ug/mL of Rabbit Anti-LC3B Polyclonal Antibody (Catalog # NB100-2220) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog# HAF008). A specific band was detected for LC3B at a molecular weight of approximately 15 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. GAPDH is shown as a loading control. This experiment was conducted under reducing conditions.

<b>Genetic Strategies Validation. </b>LC3B was detected in immersion fixed CQ treated HeLa cells (left) but was not detected in LC3B knockout HeLa cells (right) using rabbit anti-human LC3B polyclonal antibody (Catalog #NB100-2220) at 0.3 ug/mL for 3 hours at room temperature. Cells were stained using the NorthernLights(TM) 557-conjugated anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm.

<b>Biological Strategies Validation. </b>Contribution of autophagy to lipid droplet formation in WT and KO NSC34 cells.(A) WT and KO NSC 34 cells were treated with (+) or without (-) bafilomycin A (bafA) under basal or HBSS starvation conditions for 4 h and blotted for p62, LC3, and GAPDH. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32321733) licensed under a CC-BY license.

TMEM16A inhibited AngII-induced autophagy in aortas. (D) Representative immunofluorescence staining of LC3 (green) and a-SMA (red) in aortas. Scale bars, 20 um. Quantification of LC3B puncta. Five random fields (2,500 um2/filed) were measured in one section. **P < 0.01 vs. sham + TMcon; ##P < 0.01 vs. AngII + TMcon, one-way ANOVA. n = 14 sections from seven mice per group. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32226533) licensed under a CC-BY license.

LC3 accumulation in muscle fibers from patients with PAD (row B - E). Normal fibers from a Non-PAD sample (row A). Scale bar = 100 uM. Image from verified customer review.

Localization of LC3 by immunohistochemical method. The immunohistochemical staining shows that LC3, CRF, and HIF-1alpha have an overlapping localization in villous and extravillous trophoblast. d: decidua; v: villi; DeVe: decidual vessel. Original magnification 40x. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/bmri/2013/689768/) licensed under a CC-BY license.

Detection of LC3B in mouse ES cell lysates using Rabbit anti- [Catalog # NB100-2220]. LC3 detected at a molecular weight of approximately 17 kDa. Atg5-/- ES cells from Dr. Noboru Mizushima [Mizushima, N. et al. J. Cell Biol. 152 (2001)] Photo courtesy of Dr. Beth Levine, UT SW Medical Center.

ORP1L and VAP-A form cholesterol-dependent ER-AV contact sites MelJuSo cells expressing GFP or GFP-tagged ORP1L mutants were fixed and stained for LC3 and Calnexin. Y477 and D478 in the FFAT motif were mutated to alanines in the ORP1L ydaa mutant. Scale bar, 10 um. Right: co-immunoprecipitation for ORP1L (mutants) with VAP-A. GFP-ORP1L mutants or GFP were isolated from lysates of HEK293T cells co-overexpressing HA-VAPA using GFP-Trap beads. Western blot filters were probed for isolated GFP-tagged proteins, the associated HA-VAP-A and the input HA-VAP-A, as indicated. Image collected and cropped by CiteAb from the following publication (//www.nature.com/doifinder/10.1038/ncomms11808), licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>Lysates of mouse NIH3T3 and rat PC-12 cell lines untreated (-) or treated (+) with CQ. PVDF (Polyvinylidene difluoride) membrane was probed with 0.5 ug/mL rabbit anti-LC3B polyclonal Antibody (Catalog # NB100-2220, Novus Biologicals), followed by 1:2000 dilution of goat anti-rabbit IgG secondary antibody. LC3 detected at a molecular weight of approximately 15 kDa in treated NIH3T3 and PC-12 cells.

<b>Biological Strategies Validation. </b>Detection of LC3I and LC3II in mouse cochlea cell line SV-K1 using Rabbit anti- [Catalog # NB100-2220]. Cells were treated with CQ (1 uM), and As2O3 (1 uM) for 24 hrs. Image from verified customer review.

Autophagy Investigation. (A) Representative WBs (Western Blots) of LC3B-I, LC3B-II and GAPDH loading control in DANs derived from 4 iPD patients and 4 healthy controls. (B) WBs from three independent experiments were quantified by densitometry and LC3B-II normalized to a loading control. Grouping of data by healthy vs iPD as well as iPD stratified for T/T and C/C genotype. (C) Ratio of LC3B-II (normalized to loading control) following bafilomycin treatment/untreated. Citation: Marrone L, Bus C, Schondorf D, Fitzgerald JC, Kubler M, Schmid B, et al. (2018) Generation of iPSCs carrying a common LRRK2 risk allele for in vitro modeling of idiopathic Parkinson's disease. PLoS ONE 13(3): e0192497. //doi.org/10.1371/journal.pone.0192497

<b>Biological Strategies Validation. </b>Expression of autophagic protein LC3 in BME-UV1 cells forming acinar structures on Matrigel. Representative images of Western blot analysis of LC3 in bovine MECs grown in 3D culture for 3, 6, 9, and 14 days in differentiation medium (control), enriched with 17beta-estradiol (E2, 1 nM), progesterone (P4, 5 ng/mL), or both (E + P); expression of beta-actin was used as a loading control; graphs below the images show the results of densitometric analysis, in which IOD of each band was measured, and the values were normalized to IOD of beta-actin; the IOD results are presented as means +/- SEM from at least three separate experiments. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/bmri/2014/382653/) licensed under a CC-BY license.

GNA triggers the formation of autophagic markers in A549 and HeLa cell. Effects of GNA on LC3 protein. A549 cells were treated with various concentrations of GNA for 24 hours or 3 uM GNA for the indicated periods of time, then analyzed by western blotting using anti-LC3 antibodies. GAPDH protein was used as the loading control. The bar graph shows the band intensities of LC3-II relative to those of GAPDH. Mean+/- SEM, n = 3, *means p<0.05, ** p<0.01, one-way ANOVA. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0083604), licensed under a CC-BY license.

Autophagosomes accumulate in neurons of the VH gray matter at day 1 after SCI. Representative images of IHC staining for LC3 (green) and neuronal marker NeuN (red) in VH of gray matter from sham and SCI animals. Stronger co-localization between NeuN and LC3 is apparent at day 1 after SCI. Scale bar is 20 um. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2014527) licensed under a CC-BY license.

Autophagosomes accumulate in microglia and oligodendrocytes of the dorsal white matter adjacent to injury after SCI. Representative images of IHC staining for LC3 (green) and activated microglia marker CD11B (red) in dorsal white matter of sham and SCI animals. Increased co-localization between LC3 and CD11B is apparent at day 1 after SCI. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2014527) licensed under a CC-BY license.

Staining of rat brain tissue section. Analysis using Rabbit anti-LC3B Antibody [Catalog # NB100-2220]. Image from verified customer review.

Human ovarian Cancer tissue stained using heat mediated antigen retrieval in pH 6.0 citrate buffer at 1:200 dilution. Image provided by verified customer review.

FFPE (Formalin-Fixed Paraffin-Embedded) tissue section of mouse brain using 1:200 dilution of Rabbit anti-LC3B antibody [Catalog # NB100-2220]. The specific signal of LC3 was detected using HRP-conjugated secondary antibody with DAB (3, 3 -diaminobenzidine) reagent, and nuclei of cells were counterstained using hematoxylin. This LC3B antibody generated a low to moderate levels of cytoplasmic staining in the glial cells. The neurons depicted a moderate to strong staining for LC3 in their cytoplasm.

Analysis using the Biotin conjugate of Rabbit anti-LC3B Antibody [Catalog # NB100-2220]. Staining of brain, cerebral cortex, neurons with cell processes.

SCI leads to lysosomal dysfunction. Representative high magnification images ( x 60) of LC3 (green) and CTSD (red) staining in VH of gray matter from sham and day 1 SCI animals. Decrease in number and intensity of CTSD puncta (lysosomes) and increase in LC3 puncta (autophagosomes) is apparent after SCI. Scale bar is 10 um. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2014527) licensed under a CC-BY license.

Image shows a specific band for LC3B at a molecular weight of approximately 15 kDa in 0.5 mg/mL of Neuro2A lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

LILRB2 antagonism effects on the bactericidal function of the MDSCs are independent of autophagy. (B) Lysates of cells from panel A were collected at 18 h and analyzed using Western blot for the autophagy marker, microtublue-associated light chain-3 (LC3). Lipidation is indicated by LC3-II. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35757769), licensed under a CC-BY licence.

$CQ-induced autophagy inhibition leads to Nrf2 pathway activation in C2C12 muscle cells. a. Treatment of differentiated C2C12 myotubes with autophagy inhibitor CQ increased the sarcoplasmic level of LC3-II and SQSTM1 proteins in a dose- and time-dependent manner, indicating effective inhibition of the autophagic flux. Autophagy inhibition was accompanied by a slight decrease in the sarcoplasmic level of the soluble Keap1 protein (possibly reflecting its sequestration into insoluble protein aggregates) and an increase in the nuclear level of Nrf2 protein. GAPDH and lamin B were used as loading controls for sarcoplasmic and nuclear protein-enriched fractions, respectively; a representative of 3 independent experiments is shown. Image collected and cropped by CiteAb from the following publication (//actaneurocomms.biomedcentral.com/articles/10.1186/s40478-016-0384-6), licensed under a CC-BY licence.$

Accumulation of an autophagosome marker in the presynaptic terminals of NMJs of SOD1G93A mice (A) Representative confocal micrographs of immunohistochemistry showing accumulation of the autophagosome marker LC3 (grayscale in left panels, green in right panels) in presynaptic terminals of diaphragm NMJs in SOD1G93A mice at P120, but not at NMJs of wild-type mice at P140. Nerves were stained using anti-neurofilament and anti-SV2 antibodies (blue), and postsynaptic acetylcholine receptors were labeled with Alexa 594-conjugated alpha-bungarotoxin (red). In these cross-section images of NMJs, presynaptic terminals are above the bungarotoxin signal, and postsynaptic myotubes are below the bungarotoxin signal. Scale bar: 10 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28890682), licensed under a CC-BY licence.

CAIX expression in PDAC and association with outcomeA. Representative images of immunohistochemical staining showing: negative expression of CAIX in both tumor cell and stromal compartments (top left), positive expression of CAIX in tumor cells but not stromal compartment (top right), positive expression of CAIX in stromal compartment but not tumor cells (bottom left), positive expression of CAIX in both tumor cell and stromal compartments (bottom right). B. Kaplan-Meier analysis of CAIX expression in the distinct tumor cell and stromal compartments and the associated overall survival. C. Representative images of immunohistochemical staining showing low stromal volume and high stromal volume and low and high microvessel density D. Representative images of immunohistochemical staining showing: positive expression of MCT4 in tumor cell compartment and positive expression of MCT4 in tumor stromal compartment. Kaplan-Meier analysis of combinatorial MCT4 and CAIX expression in the stromal compartment and the associated overall survival. Hazard ratios and p-values for each comparison are summarized in the table. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27623078), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Al, Av, Ba, Bv, Ca, Ch, ChHa, Gp, Ha, In, Pm, Pm, Rb, SyHa, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Al, Av, Ba, Bv, Ca, Ch, ChHa, Gp, Ha, In, Pm, Pm, Rb, SyHa, Ze

Applications:

, , , , , , , , , , , ,

Applications: , , , , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-2220AF350
Alexa Fluor 405	NB100-2220AF405
Alexa Fluor 488	NB100-2220AF488
Alexa Fluor 532	NB100-2220AF532
Alexa Fluor 594	NB100-2220AF594
Alexa Fluor 647	NB100-2220AF647
Alexa Fluor 700	NB100-2220AF700
Alexa Fluor 750	NB100-2220AF750
Allophycocyanin	NB100-2220APC
Biotin	NB100-2220B
DyLight 350	NB100-2220UV
DyLight 405	NB100-2220V
DyLight 488	NB100-2220G
DyLight 550	NB100-2220R
DyLight 594	NB100-2220DL594
DyLight 650	NB100-2220C
DyLight 680	NB100-2220FR
DyLight 755	NB100-2220IR
FITC	NB100-2220F
HRP	NB100-2220H
Janelia Fluor 549	NB100-2220JF549
Janelia Fluor 646	NB100-2220JF646
PE	NB100-2220PE
PerCP	NB100-2220PCP
mFluor Violet 450 SE	NB100-2220MFV450
mFluor Violet 500 SE	NB100-2220MFV500
mFluor Violet 610 SE	NB100-2220MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-2220SS

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NB100-122

HIF-2 alpha/EPAS1 Antibody - BSA Free

(37 Reviews)

(781 Publications)

0.1 ml

NB100-122

WNT11 is induced by hypoxia or hypoxic mimetics in different cell types. Immunoblot analyses of HeLa cells under normal air or hypoxia for 24 hrs. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520) licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>HIF-1alpha is the predominant transcriptional regulator of WNT11 expression during hypoxia. EMSCs isolated from the indicated mouse genotypes were infected with lentivirus expressing GFP or Cre recombinase. Non-infected cells and GFP infected cells served as controls. Immunoblot analyses of EMSCs derived from the indicated genotypes treated with 0.1 mM DMOG for 24 hrs. Attenuated WNT11 expression in Hif-1alpha KO EMSCs (lenti-Cre infected Hif-1af/f). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520) licensed under a CC-BY license.

<b>Orthogonal Strategies Validation. </b>Formalin-fixed paraffin-embedded tissue sections of human placenta were probed for HIF-2 alpha/EPAS1 mRNA (ACD RNAScope Probe, catalog #410598; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using Rabbit Polyclonal (Novus Biologicals catalog # NB100-122) at 1:100 dilution with one-hour incubation at room temperature followed by incubation with anti-rabbit IgG VisUCyte HRP Polymer Antibody (Catalog # VC003) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to trophoblastic cells.

Expression of hypoxia-inducible alpha subunits in normal and diseased lung tissue. HIF-2alpha expression are more evident in fibroblasts from idiopathic pulmonary fibrosis (d) than from lung tissue affected by other inflammatory conditions (i.e. chronic bronchitis, panel e) or normal lung tissue (f); as demonstrated by a higher proportion of positive fibroblasts (open arrows) than negative ones (solid black arrows). Image collected and cropped by CiteAb from the following publication (//respiratory-research.biomedcentral.com/articles/10.1186/s12931-019-1100-4) licensed under a CC-BY license.

Image shows a specific band for HIF-2 alpha in 0.5 mg/mL of hypoxic HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

<b>Genetic Strategies Validation. </b>Immunoblot validation of HIF2A and PHD2 KO clones using HIF2A (#NB100-122; dilution: 1/300), PHD2 (#NB100-137; dilution: 1/500). To blot HIFs factor cells were first pre-treated for 5 h with CoCl2 300 uM before protein extraction, a condition that promotes HIF factor accumulation. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-018-06988-3) licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>HIF-2 alpha in human retinal and choroidal primary endothelia lysates using .. Image from verified customer review.

<b>Biological Strategies Validation. </b>HIF-2 alpha in MDA-MB-231 cell lysate (overexpression and endogenous samples) using . did not react to HIF-1 alpha overexpression. Image from verified customer review.

Induced WNT11 expression with tumor hypoxia and WNT11 regulates tumor growth. Bevacizumab increased expression of HIF-1alpha and HIF-2alpha and WNT11. First 10 lanes are control tumors, and the last 10 lanes are tumors from bevacizumab-treated animals. Lysates from whole tissue and nuclei are indicated. Alpha -Tubulin, actin and lamin A/C are loading controls. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520) licensed under a CC-BY license.

MDA-MB-231 cells were exposed to 20% or 1% O2 for 16 hours, and chromatin immunoprecipitation (ChIP) was performed with the indicated antibody (Ab). Primers flanking the HIF binding site were used for qPCR. ChIP image submitted by a verified customer review.

<b>Biological Strategies Validation. </b>Lane 1: Cobalt chloride treated COS7 nuclear extracts. Lane 2: Untreated COS7 nuclear extracts.

786-O cells without or with VHL overexpression. Image from verified customer review.

<b>Biological Strategies Validation. </b>Analysis using the HRP conjugate of NB100-122. Detection of normoxic and hypoxic nuclear rat cell lysates.

HIF-2 alpha immunoreactivity in human cardiac myocytes stained with NB100-122.

Expression and correlation of 3 antibodies: HIF-1 alpha, HIF-2 alpha and HAF in ccRCC. (A, C & E) Positive nuclear staining to the 3 antibodies: HIF-1 alpha, HIF-2 alpha and HAF, in primary ccRCCs at high magnification (100X). (B, D & F) Heterogenous nuclear staining of 3 antibodies in a tumor at low magnification (20X). (G, H and I). Correlation of 3 antibodies with the Fuhrman grade.Citation: Ambrosetti D, Dufies M, Dadone B, Durand M, Borchiellini D, Amiel J, et al. (2018) The two glycolytic markers GLUT1 and MCT1 correlate with tumor grade and survival in clear-cell renal cell carcinoma. PLoS ONE 13(2): e0193477. //doi.org/10.1371/journal.pone.0193477

<b>Genetic Strategies Validation. </b>Reduction of HIF-2alpha levels leads to protection in UV-triggered apoptosis, but not for apoptosis caused by glucose and serum starvation in 786-O cells. Parental 786-O or those either stably expressing wild-type VHLp19 or stably infected with a control vector (pSuperRetro) or a pool of two HIF-2alpha shRNAs vectors [21] were grown to confluence and lysed. Cell alpha-tubulin.

Sp1 and HIFs contribute to the synergistic activation of multiple genes in OVSAYO cells under SSH.b) Activation of multiple genes under hypoxia is dependent on HIFs. Western blotting is also shown for HIFs. Data shown are the mean (n = 3) ± SD.

Manipulation of DNA methylation at HRE sequences alters HIF binding. (A) The Hypoxia Response Element (HRE) contains a CpG site that can be methylated which prevents HIF1 and HIF2 binding in vitro. EMSA of in vitro-translated HIF1 and HIF2 binding to 32P-labelled unmethylated and methylated HRE probes. Competition with 250X molar excess of unlabelled/unmethylated HRE probe (lanes 3, 7,12, 16) or unlabelled control HRE-free probe (lanes 4, 8, 13, 17). HIF1 and HIF2 complexes bound to unmethylated HRE or methylated HRE supershifted with anti-HIF1alpha (lanes 9 and 18) and anti-HIF2alpha (lanes 5 and 14). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-018-21524-5), licensed under a CC-BY licence.

Hypoxia and HIF-1 accumulation after DSS-treatment.Real time PCR of HIF-1alpha exon 2 (A) in RNA samples of hypoxic treated BMDM of wild type (HIF-1alpha+f/+f) and knockout (Lyz2-Cre/HIF-1alpha+f/+f) mice with 14 samples/group. Each bar represents the mean value +/- SEM. ***P 0.01 vs. normal thyroid; #P > 0.01 vs. normal thyroid; (d) Relative expression levels of HMGB1 in thyroid cancer. Total mRNA was extracted from thyroid cancer patients’ tissues and HMGB1 level determined by relative optical intensity (in arbitrary units, AU) of bands on RT-PCR. Each dot represented relative level of HMGB1 in an individual sample. *P > 0.01 vs. FTC; (e) HMGB1 expression level for differentiating thyroid cancer tissues from non-thyroid cancer tissues in our validated cohort; AUC: 96.7%, sensitivity: 88.9% and specificity: 96.2% in the validated cohort Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31331356), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Loss of MUS81 activates DNA damage responseA. MUS81 depleted U2OS cells accumulate NBS1 foci defining sites of DNA damage. B. Partial co-localization of NBS1 foci and RPA foci in MUS81-depleted cells. C. Co-localization of RPA foci to ssDNA regions. Cells were labelled with BrdU for 3 days concomitant with MUS81 depletion. BrdU was detected at ssDNA regions by immunofluorescence. D. NBS1 foci arise in cells that are both positive and negative for Cyclin A expression. Scale bars, 5 μm. E. NBS1 foci form largely at non-telomeric loci. F. Partial co-localization of NBS1 foci with PML nuclear bodies. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/26415217), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemical detection of HIF-1 alpha protein in human tissues. The nature of the tissue is indicated on top of each figure. Original magnifications are as follows: A, ×400; B, ×400; C, ×100; D, ×100; E, ×100; F, ×100; G, ×40; H, ×40. Image collected and cropped by CiteAb from the following open publication (//bmcgenet.biomedcentral.com/articles/10.1186/1471-2156-5-27), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

ROS overproduction mediated apoptosis induced by combined treatment of DDP and HIF-1 alpha in PC-3 xenografts and cell culture. (a) ROS monitored by DHE (red) and nuclei by DAPI (blue) staining in PCa xenografts (scale bars, 50 µm). (b) MDA formation of PCa xenografts was examined after various treatments (c,d). PC-3 cells were treated with DDP, si-HIF-1 alpha plasmid, or both, in the presence or absence of NAC (5 mM) or DHLA (0.25 mM) for 24 h. Total lysates (c) and culture media (d) were used to detect cellular H2O2 level. (e,f) Western analysis for HIF-1 alpha as well as cleaved caspase-3 and PARP in PC-3 cells following various treatments. Data were presented as mean ± SD of three independent experiments. *p 2000 neurons/condition; one-way ANOVA with Bonferroni's multiple comparisons test). (B) PGD2 did not induce pRII in non-neuronal cells of the same cultures shown in A. (C) Time course of RII phosphorylation indicating long-lasting effects of PGD2 (10 µM) on sensory neurons. (D) Stimulation with PGD2 also results in phosphorylation of the ERK1/2 measured in the same cultures shown in D. (E) Representative experiment demonstrating that induction of RII phosphorylation is enhanced in TRPV1(+) neurons (total of 3664 neurons). Plots of respective controls are shown in S2 Fig. (F) Fold changes of pRII intensities in TRPV1(−) (grey bars) and TRPV1(+) (black bars) neurons after 1 min stimulation with 10 µM PGD2 (n = 3,>2000 neurons/condition, one-way ANOVA with Bonferroni's multiple comparisons test). (G) Co-labeling of TRPV1 and PTGDS revealing that PTGDS is expressed in neurons lacking TRPV1 (total of 9951 neurons, also refer to S2 Fig. for control plots). (H) Co-labeling of NF200 and PTGDS showing that PTGDS(+) neurons express NF200 (total of 12966 neurons, also refer to S2 Fig. for control plots).(I) Size distribution of PTGDS(+) (green), NF200(+) (red), and all sensory neurons (black) indicating that PTGDS(+) neurons are larger than other neurons. (J) Suggested pathway of interneuronal communication between subgroups of sensory neurons. Large-diameter mechanosensitive neurons express PTGDS resulting in the production of PGD2, which activates DP1 receptors present on nociceptive neurons expressing TRPV1. Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.pone.0115731), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

SPOC1 is recruited to I-SceI-induced DSBs where it colocalizes with 53BP1. (A) In situ immunofluorescence studies with U2OS19 ptight13 GFP-LacR cells with a stably integrated I-SceI cleavage site flanked by lac operator repeats reveals localization of the GFP-lac repressor protein (GFP-LacR) at the lac-operator DNA sequences in the nucleus before (−I-SceI) and 16 h after I-SceI-induced (+ I-SceI) DSB. In contrast, SPOC1 (white) and 53BP1 (red) are distributed throughout the nucleus in the absence of I-Scel, and partially colocalized in naturally occurring repair foci. After I-SceI cleavage to generate DSBs, SPOC1 and 53BP1 colocalize at distinct foci, including the cleaved DNA adjacent to DNA-bound GFP-LacR. Proteins were visualized by immunostaining and confocal microscopy. Scale bars = 10 μm. (B) Quantitative analysis of SPOC1 recruitment to the 53BP1 and GFP-LacR positive lacO array before and 16 h after I-SceI induction. (C) Monitoring of the kinetics of SPOC1 and 53BP1 recruitment to DSBs between 0 and 24 h post-I-SceI induction. (D) Quantifying ATM kinase inhibitor-mediated effects on recruitment of SPOC1 and 53BP1 to DSBs as evident 16 h after I-SceI-induced cleavage. (E) SPOC1 and 53BP1 colocalize at a large discrete endogenous repair focus as observed in some non-irradiated U2OS cells by immunostaining. Scale bar = 10µm. Image collected and cropped by CiteAb from the following open publication (//academic.oup.com/nar/article-lookup/doi/10.1093/nar/gks868), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Hypoxia reduces inflammatory signaling pathways and NLRP3 expression and induces autophagy in IECs. a HT-29 cells were subjected to normoxia and hypoxia at the indicated times in the absence or presence of 10 µg/ml LPS. Autophagy was measured by variations in the ratio of LC3-II/LC3-I and the total amount of LC3 (LC3-I plus LC3-II) relative to beta -actin. Results are representative of two independent experiments. b, c and d HT-29 cells were subjected to normoxia and hypoxia for the indicated periods in the absence or presence of 10 µg/ml LPS, followed by transcript analysis. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post-test. Results represent mean + s.e.m. of two independent experiments done in triplicate, *P < 0.05; **P < 0.01; ***P < 0.001; ns not significant. e HT-29 cells were subjected to normoxia or hypoxia in the presence and absence of 10 µg/ml LPS and 20 µM of MG132. Results are representative of two independent experiments. f and g Putative binding sites for HIF-1 alpha and NF-kappa B were found in the p62 f and NLRP3 g promoters using Genomatix software tools. Numbers under the boxes indicate the distance from the transcription start site. HT-29 cells were subjected to normoxia (21% O2) or hypoxia (0.2% O2) for 6 h and 24 h. ChIP analysis was performed using antibodies against HIF-1 alpha and NF-kappa B for immunoprecipitation. PCR was performed using the promoter-specific primers for the p62 f and NLRP3 g promoter binding sites of HIF-1 alpha and NF-kappa B. Aliquots taken prior to immunoprecipitation were used as input control. PCR products were run on 2% agarose gel. The results are representative of three independent experiments. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28740109), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

The FANCD2 NLS is required for the nuclear localization of a subset of FANCI.(A) FA-D2 patient cells or FA-D2 cells stably expressing FANCD2-WT were incubated in the absence (NT) or presence of MMC for 24 h, fixed, stained with rabbit polyclonal anti-FANCD2 or anti-FANCI antibody and counterstained with phalloidin and DAPI. AF-488, Alexa Fluor 488. (B) FA-D2 cells stably expressing LacZ, FANCD2-WT, FANCD2-∆N57, FANCD2-∆N100, and FANCD2-3N were incubated in the absence (NT) or presence of MMC for 24 h, fixed, and stained with rabbit polyclonal anti-FANCI antibody, and counterstained with phalloidin and DAPI. At least 300 cells were scored for cytoplasmic (Cyto.), nuclear (Nucl.), and both cytoplasmic and nuclear (Both) localization of FANCI. (C) COS-7 cells were transiently transfected with no DNA, FANCI-GFP, FANCI-GFP plus FANCD2-V5-WT, or FANCI-GFP plus FANCD2-V5-∆N57. Whole-cell lysates were immunoprecipitated with anti-V5 or anti-GFP antibodies and immune complexes immunoblotted with anti-GFP and anti-V5 antibodies. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/24278431), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Analysis of microvascular density and HIF-1 alpha activity. Microvascular density of pulmonary tumors was significantly higher in hyperplastic (B) and tumoral lesions (D) of NNK/NTHi treated mice compared to the NNK treated mice (A, and C) detected by CD105 immunostaining (10X, scale bar = 100 μm). HIF-1 alpha immunostaining after NNK/NTHi combined treatment showed hot-spots of high stromal expression in tumors (F), and high expression in perivascular-peribronchiolar lymphocytes (H). In contrast, low, homogenous expression of HIF-1 alpha was detected in the tumors (E) and perivascular-peribronchiolar lymphocytes (G) of NNK treated mice (40X, scale bar = 25 μm). Image collected and cropped by CiteAb from the following open publication (//molecular-cancer.biomedcentral.com/articles/10.1186/1476-4598-11-4), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

JMJD2B expression increases in hepatic steatotic cell and animal models. (A) HepG2 cells were incubated with a mixture of palmitic acid (PA) and oleic acid (OA) (1:2 ratio) at 800 μM concentrations for 24 h, and intracellular triglyceride (TG) levels were analyzed by a TG assay kit. JMJD2B mRNA and protein levels were examined by qPCR and western blotting, respectively. Data represent means ± SEM of three independent experiments performed in triplicate. *p < 0.05 vs. no treatment. The full-length western blots corresponding to truncated blots are presented in Supplementary Figure S1A. (B) HepG2 cells were treated with T0901317 (10 μM) for 24 h, and intracellular triglyceride (TG) levels were measured by a TG assay kit. JMJD2B mRNA and protein levels were examined by qPCR and western blotting, respectively. Data represent means ± SEM of three independent experiments performed in triplicate. *p < 0.05 vs. no treatment. The full-length western blots are presented in Supplementary Figure S1B. (C) Total RNAs were isolated from the livers of HFD-induced obese mice. The JMJD2B mRNA levels were assessed by qPCR. Data represent means ± SEM of 5 mice. *p < 0.05 vs. ND mice. ND: normal diet. HFD: high fat diet. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30214048), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Protein expressions of intrarenal RAS components in two groups of mice. (A) Immunohistochemical analysis of intrarenal RAS expression position in the two groups of mice. AGT immunoreactivity in the proximal tubular cells, Ang II immunoreactivity in both glomerular and tubular cells, renin immunoreactivity in juxtaglomerular apparatus cells, ACE immunoreactivity in brush border membranes of proximal tubules as well as AT1 and AT2 immunoreactivity in the proximal tubules were increased in the HF group when compared to the control group. (B) Western blot analysis for protein expressions of intrarenal RAS components in the two groups of mice. (C) The histogram represents mean ± SD of the densitometric scans for the protein bands of angiotensinogen (AGT), angiotensin II (Ang II), renin, angiotensin converting enzyme (ACE), angiotensin II type 1 receptor (AT1), angiotensin II type 2 receptor (AT2) from five experiments, normalized by beta -actin. * P < 0.05 vs. control. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/23570453), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$TAZ induces Tfam protein expression at the translational level.a The expression of mitochondrial biogenic factors, including Tfam, Ppargc1a (Pgc1 alpha ), Nrf1, and Nrf2, was analysed using quantitative reverse transcription (qRT)-PCR in the gastrocnemius muscle of wild-type (WT) and muscle-specific TAZ knockout (mKO) mice; n = 6 for each condition (Tfam; *p = 0.0498, Nrf1; *p = 0.0467). b Protein was analysed via immunoblotting to detect TAZ and the indicated mitochondrial biogenic factors in (a). Alpha-tubulin was used as the loading control. c RNA was isolated from the control (Con) and TAZ knockdown (KD) C2C12 myotubes, and the expression of the indicated mitochondrial biogenic factors was analysed using qRT-PCR. The experiments were performed in triplicate (Tfam; *p = 0.04). d Protein from the Con and TAZ KD C2C12 myotubes was assessed via immunoblotting to detect TAZ and mitochondrial biogenic factors. Beta-actin was used as the loading control. e RNA was isolated from WT, TAZ-knockout (KO), and TAZ-rescued (T) mouse embryonic fibroblasts (MEF), and the indicated mitochondrial biogenic factors were analysed via qRT-PCR. f Protein from WT, KO, and T MEF was assessed via immunoblotting to detect TAZ and the indicated mitochondrial biogenic factors. Beta-actin was used as the loading control. g Protein was isolated from the gastrocnemius muscle of WT and mKO mice and analysed via immunoblotting to detect TAZ, YAP, and proteins involved in the translation of both phosphorylated and total forms. Alpha-tubulin was used as the loading control. h Protein obtained from the Con and TAZ KD C2C12 myotubes was assessed via immunoblotting to observe TAZ and proteins involved in translation. Beta-actin was used as the loading control. i Protein was isolated from WT, KO, and T MEF and analysed via immunoblotting to detect TAZ and proteins involved in translation. Vinculin was used as the loading control. j The polysome profile of cells described in panel i was evaluated continuously by measuring absorbance at 254 nm. The 40S, 60S, 80S, and polysome fractions are denoted by the corresponding peaks. k The distribution of Tfam, Atp5d, and Gapdh mRNA from cells described in (j) across a density gradient was analysed using reverse transcription (RT)-PCR. bp = base pairs. For a, b, g 8 to 10-week-old mice were used. Data are presented as mean ± SEM for (a), mean ± SD for (c, e). Statistical significance was analysed via two-sided t-test for (a) and one-sided t-test for (c). One-way ANOVA with Tukey’s multiple comparison test was used for (e). Representative data was shown and experiments were performed at least twice with similar results for (b, d, f–i, k). Source data are provided as a Source data file. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35115527), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Binding of TRF2 to viral DNA during HHV-6A/B infection.A) Schematic representation of the HHV-6A/B genome. The DR6 probe used for hybridization is shown in red. Uninfected and HHV-6A-infected HSB-2 cells (B-D) or uninfected and HHV-6B-infectd Molt-3 cells (E-F) were analyzed for TRF2 binding to viral DNA using ChIP. The input was hybridized with Alu probe to assess quantity of starting material. Anti-IgG (negative control), anti-PolII (positive control) or TRF2 antibodies were used for immunoprecipitation. B) QPCR detection of GAPDH DNA following ChIP. Results are expressed as fold increase over control IgG. C and E) Eluted DNA was hybridized with 32P-labeled Alu, telomeric (TTAGGG)3 or HHV-6A (DR6) probes. After hybridization the membranes were washed and exposed to X-ray films. D and F) Densitometric analysis of relative binding of TRF2 to telomeric and viral DNA. Results of one experiment representative of three are presented and are expressed as signal after normalization to input. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32320442), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

miR-375 control on CIP2A-MYC pathway also contributes to p21 elevation. (A) p21, p53, RB, CIP2A, and MYC protein levels in MCF7 cells transfected with miR-375 inhibitor, -mimic, or NS control were measured by Western blot analysis. Tubulin expression was used as internal control. 25%, 50%, 100% amounts of untreated cell lysates were included to calibrate the semiquantitative measurement. (B) Transfection with miR-375-mimic significantly upregulated p21 mRNA in MCF7. Relative endogenous p21 mRNA levels were measured in MCF7 cells transfected with miR-375 inhibitor, -mimic, or NS control for 48 h using qRT-PCR. (C) CIP2A and MYC protein levels were effectively silenced by si-CIP2A transfection with 1 and 10 nM concentrations for 48 h. Increased p21 protein levels were detected in si-CIP2A dose-dependent manner. 10 nM of si-GFP was used as a control. (D) Protein levels of CIP2A, p53, and p21 in MCF7 cells transfected with si-p53 and/or miR-375-mimic were measured by Western blot analysis. (E) mRNA levels of p21 in MCF7 cells transfected with si-p53 and/or miR-375-mimic were measured by qRT-PCR. (F) Flow cytometry analysis demonstrates G1 arrest of MCF7 cells 48 h after transfection with miR-375-mimic compared to miR-375 inhibitor or NS control. The concentrations of siRNA or miRNA used in panels D, E, and F were 10 nM and 25 nM, respectively. (G) Schematic depiction of miR-375-mediated repression of CIP2A, E6, E6AP, and E7 in HPV16-positive cells that simultaneously increases tumor suppressor p53, p21, and RB, and causes cell cycle arrest. Results are expressed as mean ± SD from three independent experiments. *p < 0.05, **p < 0.01, and ***p < 0.001. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/24708873), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Effect of the four most common herbal formulas and single herbs on the phosphorylation of myosin light chain (MLC) protein.Briefly, A10 cells were treated with herbal formulas (A) or single herbs (B). Y27632 (Y10; 10 μM) and calyculin A (A50; 50 μg/ml) were used as negative and positive controls. Western blot analysis and staining with anti-phospho-MLC, anti-total-MLC, and anti-beta actin antibodies was then performed. Phospho-MLC, total-MLC, and beta actin were all obtained with their appropriate protein size bands. The relative Phospho-MLC intensity (%) was expressed as [(Phospho-MLC/total-MLC)drug treated/ (Phospho-MLC/total-MLC)cell only x 100%]. The Mean±SEM values for at least three independent experiments along with the representative western blot were performed. Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.pone.0145109), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Silencing CIP2A caused decreased Cdk1 and Cdk2 proteins in 16E6‐expressing cells. A, Western blot analysis of CIP2A, Cdk4, Cdk6, cyclin D1, Cdk1, Cdk2, cyclin B1, cyclin A2 and cyclin E1 protein levels in cells expressing HPV‐16E6 transfected with CIP2A siRNA and then treated with PBS or 10 μg/mL bleomycin for 24 h. A representative of 3 independent experiments is shown. B, Quantification of all cell cycle‐related proteins. Data from 3 experiments are summarized. C, Relative mRNA levels of all cell cycle‐related genes determined by qRT‐PCR. Data from 3 experiments are summarized. *, P < .05; **, P < .01 Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29893470), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Effect of alcohol on TERT expression in liver tissues and KCs during ALD development.(a) TERT expression in liver tissues was performed by IHC analysis. Representative views from each group were presented (original magnification, ×40). (b) Total TERT mRNA and protein levels in liver tissue were analyzed by real-time PCR and western blot. The results are shown as relative expression against control expression without treatment. (c) Representative colocalization of TERT with macrophage CD68 immunoreactivity in liver tissue by using the double immunofluorescent (IF) analysis. (d) Total TERT mRNA and protein levels in KCs isolated from the liver were analyzed by real-time PCR and western blot. The results are shown as relative expression against control expression without treatment. (e) Quantification of telomerase activity (TA) in CD-fed mice and EtOH-fed mice. RNase treatment or heat inactivation of KCs isolated from the liver of EtOH-fed mice served as negative controls for the TA assay. All quantitative data are presented as mean ± SD percentage increase compared with CD-fed group (n = 4 in CD-fed group, n = 6 in EtOH-fed group) *P < 0.05, **P < 0.01 vs CD-fed group. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/26725521), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Fi, Ha, Pm, Rb, Re, Sh

Species Glossary

Reactivity: Hu, Mu, Rt, Fi, Ha, Pm, Rb, Re, Sh

Applications:

, , , , , , , , , , , , , ,

Applications: , , , , , , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-122AF350
Alexa Fluor 405	NB100-122AF405
Alexa Fluor 488	NB100-122AF488
Alexa Fluor 532	NB100-122AF532
Alexa Fluor 594	NB100-122AF594
Alexa Fluor 647	NB100-122AF647
Alexa Fluor 700	NB100-122AF700
Alexa Fluor 750	NB100-122AF750
Allophycocyanin	NB100-122APC
Allophycocyanin/Cy7	NB100-122APCCY7
Biotin	NB100-122B
DyLight 350	NB100-122UV
DyLight 405	NB100-122V
DyLight 488	NB100-122G
DyLight 550	NB100-122R
DyLight 594	NB100-122DL594
DyLight 650	NB100-122C
DyLight 680	NB100-122FR
DyLight 755	NB100-122IR
FITC	NB100-122F
HRP	NB100-122H
Janelia Fluor 549	NB100-122JF549
Janelia Fluor 646	NB100-122JF646
PE	NB100-122PE
PE/Atto594	NB100-122PEATT594
PE/Cy5.5	NB100-122PECY55
PE/Cy7	NB100-122PECY7
PerCP	NB100-122PCP
mFluor Violet 450 SE	NB100-122MFV450
mFluor Violet 500 SE	NB100-122MFV500
mFluor Violet 610 SE	NB100-122MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-122SS

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NB100-479

HIF-1 alpha Antibody - BSA Free

(28 Reviews)

(497 Publications)

0.1 ml

NB100-479

ChIP-qPCR assay of HIF-1 binding to hypoxia response element (HRE) of the PKM2 gene in human MDA-MB-231 breast cancer cells treated with vehicle or DMOG (HIF inducer) for 8 hours. Negative control: immunoprecipitation was performed with IgG. Image courtesy of Gregg Semenza, Johns Hopkins University School of Medicine.

<b>Biological Strategies Validation. </b>Normoxic pre-treatment sensitizes glioma cells to radiation after graded chronic hypoxic (GCH) exposure. The results of anchorage-independent colony forming assays are shown for U87, U87-luc, GL261 glioma cells and 0308 GSCs after 5 Gy radiation exposure under varying O2 conditions. For easier comparisons among cell types, raw values are expressed as a % of the cell type's negative (non-irradiated) control and the means & SEMs are plotted. Each result represents at least 3 independent samples, plated in triplicate. Holm-Sidak comparisons for multiple groups were used for statistical comparisons of raw values (*p0.05. (d) IDH1R132H expression diminishes DEPTOR levels. Flag-IDH1R132H was introduced in p53−/− MEFs by retroviral transduction and whole-cell lysates immunoblotted. (e) Inhibition of endogenous mutated IDH1R132C decreases mTOR activity in a KDM4A-dependent manner. HT1080 cells were transduced with lentivirus expressing sh_KDM4A.3 or sh_Control. The cells were treated with 3 μM AGI-5198 twice every day for 5 days and whole-cell lysates blotted for markers of mTOR activation. (f) Expression of mutant IDH1 in NHA-hTERT increases mTOR signalling. IDH1WT-Flag or IDH1R132H genes were introduced in astrocytes through retroviral infection and cell lysates were blotted for mTORC2 activation. (g) IDH1R132H overexpression increases 2HG production in astrocytes. Intracellular 2HG levels were normalized to glutamate. Asterisk denotes a statistical difference between IDH1R132H and empty vector cells, two-sided t-test P<0.0001 (graph represents three independent experiments). Error bars represent standard deviation. (h) Treatment of NHA-hTERT with octyl-2HG activates mTOR signalling. The cells were treated for 8 h with 1 mM octyl-2HG. The experiment shown is a representative of triplicates. (i) Non-canonical mTOR activation in IDH1/2WT and IDH1R132H human astrocytomas. Scale bar, 50 μm. Representative samples from IDH1/2WT (n=3) and IDH1R132H (n=3) human astrocytomas are shown. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/ncomms12700), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Comparison between primary tumor and mLNs of the mouse model. Expression of CA9 and GGT1 in mLNs was higher than that in the primary tumor at 1week after injection of cells. (A) CA9 and GGT1 expression of HT29 mLNs compared to primary tumor at 6 weeks or 1 week after injection of cells. (B) Tissue oxygen concentration (%) of rectal submucosa (SM) and nLNs (n = 3). Error bars represent SD. mNs: metastatic lymph nodes, nLNs: non metastatic lymph nodes. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30542087), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Hif1 alpha expression in neonatal and adult testis.(A) Section of testis from 5-day old (P5) male new born pups showing Hif1 alpha expression in MVH+ gonocytes within the seminiferous tubules (top) and negative control images without primary antibodies (bottom). Scale bar: 50μm. (B) Western blot analysis of Hif1 alpha expression in P5 testes (left) compared to extract of the adult brain sub-ventricular zone (SVZ) (right). Loading control ( beta -Actin) is shown below. (C) Section of adult (3 month old) testis showing Hif1 alpha expression in spermatogonia. Scale bar: 30μm. (D) Western blot analysis of whole adult testis. HEK293 cells treated with DFX were used as a positive control and intestinal tissue was used as a negative control. Loading control ( beta -Actin) is shown below. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27148974), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Subcellular localization of mPARM-1 and hPARM-1 (full-length and mutant proteins). (a) NIH/3T3 cells, transiently transfected with GFP-tagged mParm-1 or hParm-1 constructs were visualized using confocal microscopy. (b) For the Golgi co-localization, transfected NIH/3T3 cells were fixed and stained with Bodipy ceramide marker. For late and early endosomes co-localization, fixed cells were labeled with (c) anti-Rab5 (early endosomes (1:100, Cell signaling)) and (d) anti-Rab7 (late endosomes (1:100, Cell signaling)) antibodies, respectively. Jurkat T cells, transiently transfected with hParm-1-GFP proteins were visualized (e) without fixation or (f) following fixation and staining with CellMask plasma membrane labeling. NIH/3T3 cells were transiently transfected with (g) ∆EC-GFP, (h) ∆SP-GFP, (i) EC-GFP, (j) ∆TM-GFP and (k) ∆CT-GFP constructs of hPARM-1 and visualized using confocal microscopy. (l) hPARM-1-GFP co-localizes with microtubules. NIH/3T3 cells transfected with hParm-1-GFP construct were fixed and stained with anti-alpha -tubulin (1/2000, Sigma) antibody. Arrows indicate co-localized hPARM-1-GFP vesicles and microtubules. NIH/3T3 cells transiently expressing (m) hPARM-1-GFP or (n) ∆CT-GFP were fixed, immunostained for caveolin-1 (1:100, Novus Biologicals), and examined by confocal fluorescence microscopy. For hPARM-1-GFP-caveolin-1 co-localization, cells that clearly demonstrated cell membrane PARM-1 localization were chosen. All co-localizations were observed following merging images of GFP-tagged proteins with those of Golgi, endosomes, plasma membrane, alpha -tubulin or caveolin-1 labeling. Cells were imaged with a laser-scanning confocal microscope (Bio-Rad MRC-1024 ES) mounted on a Nikon TE-300 using a Plan Apochromat 60x (NA 1.40) oil objective (Nikon), digitally acquired using Laser Sharp software Version 3.2 (Bio-Rad). For live cell imaging, signals were collected at a rate of 2 seconds. Images were analyzed using NIH ImageJ Version 1.42l software. Data are representative of 3 independent experiments. Image collected and cropped by CiteAb from the following open publication (//molecular-cancer.biomedcentral.com/articles/10.1186/1476-4598-12-84), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Inhibition of Cdk1 and Cdk2 by CIP2A knockdown in cervical cancer SiHa cells caused G1 arrest. A, Western blot analysis of 16E6, p53 and CIP2A after HPV‐16E6 knockdown in cervical cancer SiHa cells. B, Protein expression of CIP2A, B‐Myb, Cdk1 and Cdk2 in SiHa cells after CIP2A knockdown. Data from a representative of 3 experiments are shown. C, Flow cytometric analysis of SiHa cells with CIP2A knockdown treated with PBS or bleomycin. A representative flow cytometry of 3 independent experiments is shown. D, Quantification of percentages G1 phase cells. Data from 3 experiments are summarized. *, p<0.05 Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29893470), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Immunoblot analysis of livers from control and L-Chrebp−/− mice subjected to fasting and refeeding with a high-sucrose diet. Littermate control and L-Chrebp−/− mice (same as those described in supplemental Tables S2A and S2B) were subjected to fasting and refeeding. The nonfasted (N) groups were fed chow diet ad libitum. The fasted (F) group was fasted 12 h, and the refed (R) group was fasted for 12 h and then refed with 60% (w/w) high-sucrose diet for 12 h prior to study. Liver whole-cell lysates and membrane fractions were prepared individually, and equal amounts of protein from each mouse of the same group (four per group) were pooled. Aliquots (40 μg for whole-cell lysates and 30 μg for membrane fractions) of the pooled protein were subjected to SDS-PAGE and immunoblot analysis. Immunoblot analysis of Insig-1 and Insig-2 were carried out using membrane fractions. Whole-cell lysates were used to detect other proteins. The precursor and nuclear forms of SREBPs are denoted as P and N, respectively. Calnexin was used as loading control. Image collected and cropped by CiteAb from the following open publication (//linkinghub.elsevier.com/retrieve/pii/S0022227520331369), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Reactivity:

Hu, Mu, Rt, Po, Ca, Fi, Gt, Ha, Pm, Rb, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ca, Fi, Gt, Ha, Pm, Rb, Ze

Applications:

, , , , , , , ,

Applications: , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-479AF350
Alexa Fluor 405	NB100-479AF405
Alexa Fluor 488	NB100-479AF488
Alexa Fluor 532	NB100-479AF532
Alexa Fluor 594	NB100-479AF594
Alexa Fluor 647	NB100-479AF647
Alexa Fluor 700	NB100-479AF700
Alexa Fluor 750	NB100-479AF750
Allophycocyanin	NB100-479APC
Allophycocyanin/Cy7	NB100-479APCCY7
Biotin	NB100-479B
DyLight 350	NB100-479UV
DyLight 405	NB100-479V
DyLight 488	NB100-479G
DyLight 550	NB100-479R
DyLight 594	NB100-479DL594
DyLight 650	NB100-479C
DyLight 680	NB100-479FR
DyLight 755	NB100-479IR
FITC	NB100-479F
HRP	NB100-479H
Janelia Fluor 549	NB100-479JF549
Janelia Fluor 646	NB100-479JF646
PE	NB100-479PE
PE/Atto594	NB100-479PEATT594
PE/Cy5.5	NB100-479PECY55
PE/Cy7	NB100-479PECY7
PerCP	NB100-479PCP
mFluor Violet 450 SE	NB100-479MFV450
mFluor Violet 500 SE	NB100-479MFV500
mFluor Violet 610 SE	NB100-479MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-479SS

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NB400-105

ABCA1 Antibody - BSA Free

(7 Reviews)

(398 Publications)

0.1 ml

NB400-105

Formalin-fixed paraffin-embedded tissue sections of human prostate cancer were probed for ABCA1 mRNA (ACD RNAScope Probe, catalog # 432291; Fast Red chromogen, ACD catalog # 322360). Adjacent tissue section was processed for immunohistochemistry using rabbit polyclonal (Novus Biologicals catalog # NB400-105) at 1.5ug/mL with overnight incubation at 4 degrees Celsius followed by incubation with anti-rabbit IgG VisUCyte HRP Polymer Antibody (Catalog # VC003) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to glandular cells.

<b>Genetic Strategies Validation and Orthogonal Strategies Validation.</b>HSKMCs were transfected with scrambled siRNA (scr siRNA) or ABCA1 siRNA. The cells were lysed and ABCA1 (A) protein levels were quantified by immunoblotting. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-018-38014-3) licensed under under a CC-BY license.

<b>Biological Strategies Validation. </b>Analysis of ABCA1 in total cell lysates of RAW264.9 cells treated with vehicle (-) or 9-cisretinoic acid and 22Rhydroxycholesterol (+). Samples used for this testing were 40 ug of total cell post-nuclear lysate from each group.

<b>Biological Strategies Validation. </b>HepG2 cells were grown to 60% confluency, serum starved for 24 hours, and then treated with 1uM TO9 for 24 hours prior to being fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-ABCA1 at 5.0ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Untreated HepG2 cells were grown to 60% confluency, and serum starved for 24 hours prior to being fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-ABCA1 at 5.0ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

ABCA-1 FITC conjugated antibody of human adipose tissue macrophage subsets by flow cytometry. Image from verified customer review.

Association between expression of ABCA1 and survival in ovarian cancer patients. Expression of ABCA1 in 55 ovarian cancer patient samples was determined by IHC in tissue microarray. Representative image of ovarian cancer showing high (left panel) and low (right panel) ABCA1 expression on the cell membrane or cytoplasm (x400). Image collected and cropped by CiteAb from the following publication (//www.clinicalepigeneticsjournal.com/content/7/1/1), licensed under a CC-BY license.

ABCA1 in human primary fibroblasts in culture. 10 ug of total protein. 7.5% TGX gel. Blocking 5% milk in PBST 1h RT. 1:1000 primary ab in BSA 3% PBST O/N at 4C. Secondary 1:5000 HRP 1h RT. Arrow shows around 250 kDa. WB image submitted by a verified customer review.

ABCA1 antibody was tested at 1: 400 in HeLa cells using an Alexa Fluor 488 secondary (shown in purple). M1 is defined by unstained cells.

<b>Biological Strategies Validation. </b>Detection of ABCA1 in mouse peritoneal macrophages using NB 400-105 (Lot L). ECL exposure, 1 min. Lane 4: T09 uninduced lysate Lane 5: T09 induced lysate.

Prostates of LXR mutant mice accumulate cholesterol esters through inappropriate LXR target genes regulation. Total protein lysates of WT and LXR null mice under normal or high cholesterol diet were analyzed by western blotting with antibodies against ABCA1, LDLR and ACTIN as a loading control (left panel), quantification of ABCA1 and LDLR protein accumulation levels (right panel). * p<0.05, *** p<0.001 in Student's t test. Error bars represent the +/- mean SEM. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pgen.1003483), licensed under a CC-BY license.

miR-33b reduces cellular cholesterol efflux and serum HDL-C levels. Western blotting for ABCA1 (using NB400-105) and ABCG1 proteins in peritoneal macrophages from WT and KI+/+ mice. Representative images are shown. beta-actin was used as the loading control. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep05312), licensed under a CC-BY license.

miR-33b regulates ABCA1 and SREBP-1. Analysis for ABCA1, SREBP-1, CPT1a, and AMPK alpha protein levels in the livers of WT, KI+/-, and KI+/+ mice. Representative images are shown. TF2B and beta-actin were used as loading controls. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep05312), licensed under a CC-BY license.

Detection of ABCA1 in human prostate epithelium showing luminal and membrane staining.

Prostates of LXR mutant mice accumulate cholesterol esters through inappropriate LXR target genes regulation.(A) Plasma concentrations of cholesterol were determined (N = 9/13 per group) after 5 weeks dietary conditional exposure in each genotype. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23675307), licensed under a CC-BY licence.

Gas6 treatment enhances expression of LXRalpha and LXRbeta and their target genes in BMDM.Mouse BMDM were stimulated with 400 ng/ml Gas6, 1 uM T0901317, 10 ng/ml interferon (IFN)-gamma, 10 ng/ml IL-4, or 100 ng/ml LPS for 4 h (a,b) or 400 ng/ml Gas6 for the indicated times (c-g). (a,b,e) The amounts of the LXRalpha, LXRbeta, ABCA1, ABCG1, ApoE, AIM, Arg2, VEGF, YM1, and Arg1 mRNAs were analyzed by real-time PCR and normalized to that of Hprt mRNA. (c,d,f,g) The relative abundances of LXRalpha, LXRbeta, ABCA1, ABCG1, ApoE, Aim, Arg2, and VEGF proteins were determined by Western blotting analysis. The relative densitometric intensity was determined for each band and normalized to beta-actin. Data in all bar graphs are means +/- SEM of three independent experiments. *P < 0.05 compared with control. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27406916), licensed under a CC-BY licence.

Western Blotting for ABCA1, as detailed in Supplementary Protocol #5 (Additional file 1). Caco-2 cells cultivated on filter inserts were treated for 48 h with bexarotene (RXR agonist, 5 uM), T0901317 (LXR agonist, 10 uM), GW3965 (LXR agonist, 10 uM) or the vehicle control (DMSO 0.1%). Protein expression was then determined by western blot analysis. Results were normalized for interexperimental variance and then set in relation to the vehicle control. Bar graphs represent mean +/- SD from three independent experiments. A representative blot is shown - the bands stem from the same membrane and the vertical line indicates excised irrelevant bands. ns not significant, *P < 0.05, **P < 0.01 versus solvent vehicle (determined by paired t-test) Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32308567), licensed under a CC-BY licence.

Anti-ApoA-1 IgGs upregulate ABCA1. ABCA1 was increased after 16 and 24 h of anti-ApoA-1 IgG exposure to HMDM at the mRNA level as revealed by RT-PCR (a) as well as at the protein level after 24 h anti-apoA-1 IgG stimulation, as revealed by Western blot analysis (b). In panel a, data are expressed as fold change expression of the mean +/- SD of ABCA1 calculated by deltadeltaCT method of six independent experiments (n = 6) and values were normalized to untreated condition. p-values were calculated using the Student's t-test: * p < 0.05, ** p < 0.01, **** p < 0.0001. (b) One of four representative Western blots is shown. Data are the mean +/- SD of band intensity volume/actin intensity volume of four independent experiments (n = 4). p-values were calculated using the Student's t-test: * p = 0.02, ** p = 0.005. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31766415), licensed under a CC-BY licence.

ApoA-I increases glucose uptake in HSKMCs in an ABCA1- and SR-B1-dependent manner. HSKMCs were transfected with scrambled siRNA (scr siRNA), ABCA1 siRNA or SR-B1 siRNA. The cells were lysed and ABCA1 (A) and SR-B1 (B) protein levels were quantified by immunoblotting. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30718702), licensed under a CC-BY licence.

Functional miR-33 responsive elements in the 3'UTR of ATP8B1 and ABCB11. G). Relative protein levels in HuH7 cells transduced with empty or miR-33 adenovirus. Some cells were incubated for 16 h in the presence of FXR:RXR agonists (2 umol/L GW4064 : 1 umol/L 9-cis-retinoic acid) to induce ABCB11. Asterisk indicates a non-specific band. Data shown as mean +/- SD; **p < 0.01. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/22767443), licensed under a CC-BY licence.

LXRs control expression of genes involved in cholesterol homeostasis and fatty acid synthesis in MPECs. (D) Western blot analysis was performed on WT (+/+) or Lxralphabeta-/- MPECs using Srebp1c, Abca1 and beta-Actin antibodies. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23554947), licensed under a CC-BY licence.

Suppressive effects of Ucn1 on foam cell formation and related protein expression in human monocyte-derived macrophages.Human monocytes were incubated for 7 days with RPMI-1640 supplemented with 10% human serum and the indicated concentrations of Ucn1, followed by a 19 h-incubation with 50 ug/ml oxLDL in the presence of 0.1 mmol/l [3H]oleate. Intracellular CE accumulation was determined from the radioactivity of cholesterol-[3H]oleate. Otherwise, before the addition of oxLDL, cells were harvested and subjected to immunoblotting analyses for CD36, ACAT1, or ABCA1. beta-Actin served as a loading control. Data are expressed as means +/- SEM from 4-6 independent experiments with monocytes from 4-6 different donors. Baseline (1 fold) = 8.4+/-2.2 nmol/mg cell protein. *P<0.05, +P<0.001 vs. 0 nmol/l of Ucn1. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0110866), licensed under a CC-BY licence.

Alternatively activated macrophages (M2) exhibit increased clearance of OxLDL in vitro C). Increased ABCA1 and ABCG1 expression in M2-derived foam cells. Shown is a representative Western blot for the presence of ABCA1, ABCG1 and beta-actin in lysates of cells that were treated as indicated. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23027612), licensed under a CC-BY licence.

$Impairment of mitophagy upon Antimycin A and Oligomycin exposure.a Cells were treated with Antimycin A (1 μM) and Oligomycin (1 μM) for 1, 3, and 6 and mitochondrion/cytosol fractionation was performed to assay the accumulation of LC3-II, p62 and PINK1 within the mitochondrial fraction. VDAC1 was used as a loading control for mitochondrial fractions. b, c Cells were treated with Antimycin A (1 μM) and Oligomycin (1 μM) for 1, 3, 6, and 24 h and total fibroblasts extracts were analyzed for PINK1, p62, COXIV and VDAC1. Each quantitative data was normalized with ACTIN (n ≥ 3). Statistical analysis was performed using two-way ANOVA with Tukey’s multiple comparisons test. (*p < 0.05; **p < 0.01; ***p < 0.001). Immunoblots reported are from one representative experiment Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31332166), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

GPR50 competes with FKBP12 for the binding to T beta RI. a (Left) HEK293T cells were transfected with Flag-T beta RI alone or cotransfected with HA-T beta RII (with TGF beta ; 2 ng/mL; 1 h), myc-FKBP12 and either GPR50 delta 4 or GPR50wt. Lysates were precipitated for FKBP12 using anti-myc antibody and blotted with an anti-Flag to reveal complex formation. Expression of myc-FKBP12, Flag-T beta RI, HA-T beta RII and GPR50 was determined in total lysates. (Right) Densitometric analysis of 3 independent experiments (Mean ± s.e.m., n = 3 independent experiments, **p < 0.01, one-way ANOVA with Dunnett’s post hoc test). b, c Competition between FKBP12 and GPR50 for T beta RI binding was checked by precipitating FKBP12 from total brain and lung lysates of wt and GPR50ko mice and revealed with anti-T beta RI. Total lysate was addressed for expression of FKBP12, T beta RI, and GPR50 with corresponding antibodies. d To address the competition of FKBP12 and GPR50 for T beta RI binding, BRET measurements were performed with HEK293T cells expressing fixed amounts of T beta RI-Rluc8 and GPR50 delta 4-YFP or GPR50wt-YFP or T beta RII-YFP and stimulated with TGF beta (0.6 nM, 30 min, 37 °C). Immunoblot on the top shows FBP12 expression when transfected either with empty (Mock) or FKBP12 in different conditions. e HEK293T cells expressing the indicated proteins were starved and treated for 1 h with 2 ng/ml of TGF beta or 100 ng/ml of FK506. Total lysates were immunoblotted for Smad3 phosphorylation and total expression of myc-Smad3, GPR50 and myc-FKBP12 with suitable antibodies. f Alignment of FKBP12 and GPR50 sequences revealed similarities between the C-terminal “ATGHP” motif in FKBP12 and four repetitive motifs in GPR50 (upper top panel). One motif of GPR50 is located close to the delta 4 deletion of GPR50 delta 4 (lower bottom panel). Structural data with permission adapted from Huse et al.29 highlight the implication of the HP loop (red) in binding to T beta RI (lower panel). g HEK293T cells were transfected with indicated plasmids and as in a. Lysates were precipitated for FKBP12 using an anti-myc antibody and blotted with an anti-T beta RI to reveal complex formation. Expression of myc-FKBP12, HA-T beta RI, and GPR50 was determined in total lysates. Representative results are shown for b, c, e, and g. Similar results were obtained in at least two additional experiments. See also Supplementary Fig. 3 Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/s41467-018-03609-x), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Involvement of CBP in BNIP3 expression regulated by HIF-1 alpha and FOXO3 under hypoxia. (A) UCB-hMSCs were incubated with hypoxia condition for 24 h. Co-immunoprecipitation of HIF-1 alpha and FOXO3 with IgG and CBP were shown in left panel. IgG was used as a negative control. The total protein expressions of HIF-1 alpha , FOXO3, CBP and beta -actin in lysate were shown in right panel. n = 3. (B) CBP (20 μM) was pretreated to UCB-hMSCs, and cells were incubated with hypoxia for 24 h. The BNIP3 mRNA expression level was analyzed by qPCR. n = 6. (C) BNIP3 and beta -actin protein expressions were analyzed by western blot. Data represent mean ± S.E. n = 4. (D, E) Sample DNA was immuno-precipitated with RNA polymerase, IgG, HIF-1 alpha and FOXO3 specific antibodies. CHIP (top panel) and lysate (bottom panel) samples were amplified with the primers of GAPDH and BNIP3 promoters. Quantitative CHIP data was analyzed by qPCR, and shown in the right panel. n = 4. Western blot data were normalized by beta -actin, and qPCR data were normalized by ACTB mRNA expression level. Quantitative data are presented as a mean ± S.E.M. All blot images are representative. *p < 0.05 versus control, #p < 0.05 versus hypoxia. Image collected and cropped by CiteAb from the following open publication (//linkinghub.elsevier.com/retrieve/pii/S2213231717303804), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

DNA methylation and expression level of the PHD3 gene in HCT116 and DLD-1 CRC cells. A. HCT116 and DLD-1 cells were cultured under normoxic or hypoxic (1% O2) conditions for 48 hrs. Cells were then used for DNA isolation followed by bisulfite modification. Methylation percentage of three DNA fragments within the PHD3 CpG island (Additional file 1, Additional file 2) in HCT116 and DLD-1 cells under hypoxic and normoxic conditions was determined by Real Time PCR amplification of bisulfite treated standard and cell line DNA, followed by comparison of their HRM profiles. B. Cells were cultured in DMEM either in hypoxic (1%O2) or normoxic conditions for 48 hrs. After incubation, the cells were used for total RNA isolation and reverse transcription. The PHD3 cDNA levels were determined by RQ-PCR relative quantification analysis. RQ-PCR results were standardized by the geometric mean of PBGD and hMRPL19 cDNA levels. PHD3 cDNA levels are expressed as a multiplicity of these cDNA copies in the cell line’s calibrator. C. Cells were cultured in DMEM either in hypoxic (1%O2) (H) or normoxic (N) conditions for 48 hrs. Cells were then used for protein isolation. Proteins were separated by 10% SDS-PAGE, and transferred to a membrane that was then immunoblotted with Rp anti - PHD3 Ab and incubated with goat anti-rabbit HRP-conjugated Ab. The membrane was then stripped and reblotted with Rp anti-GAPDH Ab, followed by incubation with goat anti-rabbit HRP-conjugated Ab. The band densitometry readings were normalized to GAPDH loading control. The ratio of PHD3 to GAPDH for DLD-1 in normoxic conditions was assumed to be 1. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/24195777), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

The role of HIF-1 in purinosome formation.a, quantifying the number of purinosome-containing cells in normoxia or hypoxia (24 h) in purine-rich medium and normoxia in purine-depleted medium (purine -ve), cells in hypoxia transfected with siRNA to HIF-1 alpha (+ siRNA), and cells in purine-rich medium supplemented with DFX. Data shown are n = 3, mean ± S.E., total number of cells counted are shown in parentheses. b, time course of purinosome formation in hypoxia shows the number of purinosome-containing cells steadily increases after 3 h in hypoxia. Re-oxygenation of the samples after hypoxic incubation for 10 h reverts the number purinosome-containing cells back to normoxic levels. Data shown is n = 3, mean ± S.E., total number of cells counted are shown in parentheses. c, time course of HIF-1 alpha stabilization in hypoxia shows maximum HIF-1 alpha protein expression levels at 3 h in hypoxia, after which the HIF-1 alpha expression decreases. The positions of molecular markers are shown for each blot; uncropped blots with overlaid markers are deposited in the raw data files. d, the effect of hypoxia on the transcription of purine biosynthesis enzymes measured by qPCR. Vascular endothelial growth factor (VEGF) and HIF-1 alpha are controls. Data shown are n = 5, mean ± S.E. e, the effect of hypoxia on the protein expression levels of the purine biosynthetic enzymes. HIF-1 alpha is stabilized in hypoxia as expected, and no significant increase in the purine enzymes was detected between normoxic (21% oxygen) and hypoxic (1% oxygen) growth conditions. The positions of molecular markers surrounding each band of interest are shown for each blot; uncropped blots with overlaid markers are deposited in the raw data files. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32439803), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

FIH exits the nucleus via a Leptomycin B-sensitive exportin1-dependent pathway. (A) Immunofluorescence staining of FIH (green) and HIF1 alpha (red) in MCF7 cells after the indicated hypoxia (0.5% O2) and re-oxygenation treatments. TO-PRO-3 (blue) was used to stain nuclei. (B) Total cell lysates from U2OS cells were immunoprecipitated with an anti-exportin1 antibody or control IgG. FIH, exportin1 and beta -tubulin levels are indicated. (C) Immunofluorescence staining of FIH (green) in FIH-null mouse embryonic fibroblasts (MEFs) transfected with HA-FIH 1–349 or HA-FIH delta NES followed by normoxia, hypoxia (1% O2, 3 h) or 3 h of hypoxia followed by re-oxygenation for 1 h. TO-PRO-3 (blue) was used to stain nuclei. Arrows indicate nuclear localization of signal. (D) Total cell lysates from U2OS cells transfected with control vector, HA-FIH 1–349 or HA-FIH delta NES were immunoprecipitated with an anti-exportin 1 antibody. HA-FIH, exportin1 and beta -tubulin levels are indicated. FL, full length; IgGL, IgG light chain. Scale bars: 20 µm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30333145), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Munc13-4 KD strongly impairs exosome release. (A) SDS-PAGE Western blot of indicated proteins in MDA-MB-231 cells after stable expression of shRNA for Munc13-4 or Rab27a or a scrambled control (Ctrl). (B) Culture medium from MDA-MB-231 cells either untreated or stimulated with 1.25 µM ionomycin for 30 min was centrifuged at 1,000 g to remove cellular debris and 10,000 g to remove large extracellular vesicles. (C) The resulting 10,000-g supernatant was filtered onto a nitrocellulose membrane and analyzed for CD63, CD9, ALIX, and GM130 content by antibody blotting. (D) Quantification of CD63, CD9, and ALIX blots in C are shown as exosome release as a percentage of total cellular material with mean values ± standard error (SE) for n ≥ 3. *, P < 0.05 for comparison with corresponding control samples. (E) Panc-1 or A549 cells were left untreated or were treated with TGF beta -1 for 24 h. Indicated proteins were detected by SDS-PAGE Western blot. (F) Panc-1 cells were left untreated or were treated with TGF beta -1 for 24 h, and Munc13-4 levels were determined by immunofluorescence. TGF beta -1–treated cells exhibited a mesenchymal morphology. Bars, 5 μm. (G) A549 cells stably expressing control shRNA (Ctrl) or Munc13-4 shRNA were left untreated (Untr) or were treated with TGF beta -1 for 24 h, and SDS-PAGE Western blotting for indicated proteins was conducted. (H) Culture media supernatants (as in B) from A549 cells that were either untreated or were stimulated with 1.25 µM ionomycin for 30 min were filtered onto nitrocellulose membrane and analyzed for CD63 and GM130. (I) Quantification of CD63+ exosome release shown as a percentage of total cellular material with mean values ± SE for n = 5. *, P < 0.05; **, P < 0.01 for comparison with corresponding control samples. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29930202), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Elevated Spy1 levels prevent contact inhibition. (A) Representative views of focus formation assays in NIH3T3 cells transfected with Myc-Spy1-WT, Myc-Spy1-TST, positive control Ras-V12 or empty vector PCS3 control. n = 3 (B) Quantification of the number of colonies. Error bars reflect SE between triplicate experiments. t test was performed;** P ≤ 0.01. n = 3 (right panel) Western blot analysis of one representative lysate from A-B. Image collected and cropped by CiteAb from the following open publication (//bmccancer.biomedcentral.com/articles/10.1186/1471-2407-12-45), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

SOD2 mRNA and protein expression. RT-PCR: (A) SOD2 mRNA expression is higher in adults, and highest in the adult parenchyma. (B) No exposure effects on SOD2 mRNA were observed in neonates. (C) Adult SOD2 mRNA was decreased in PFP48. Data are presented as mean+SEM (n=5-7 rats/group, in each compartment), * significantly different compared to neonates in the same compartment, † significantly different compared to airways in the same age, ‡ significantly different compared to FA in the same compartment. Western blotting: (D) Scan of representative SOD2 and actin blots. (E) Neonatal whole lung SOD2 protein expression was unchanged with exposure, and (F) adult whole lung SOD2 protein trended upwards at PFP2, but was statistically insignificant. (G-J) Immunohistochemical localization of SOD2 in lung (n=6 rats/group). SOD2 protein was more abundant in adults compared to neonates, but no exposure specific differences were observed. Scale bar is 50 μm. Image collected and cropped by CiteAb from the following open publication (//particleandfibretoxicology.biomedcentral.com/articles/10.1186/1743-8977-10-34), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Cells of human meninges co-express LLEC markers. a–c DAB-IHC with single antibodies detects VEGFR3 (a), LYVE1 (b), and MRC1 (c) in the meninges of human post mortem brain showing no signs of neuropathology. These images are taken from a 38 year old male (sample P17/07, Table 1), and confirmed in n = 2 additional samples. P parenchyma. Scale = 150 µm (a); 40 µm (b); and 20 µm (c). d–f DAB-IHC with single antibodies detects VEGFR3 (b), LYVE1 (c), and MRC1 (d) in elderly human meninges (age: 89–92) with evidence of neuropathology and confirmed in n = 3 brains (Table 1). P, parenchyma. Scale = 20 µm. g–p IHC with fluorescent antibodies detects human meningeal cells that co-express MRC1 (h, m, yellow), LYVE1 (i, n, white), and VEGFR3 (j, o, green). Nuclei/RNA are labelled with DAPI (g, l, blue) and images are merged in (k, p). Scale = 10 µm Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31696318), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Inhibition of KDM expression in vivo.(A) Ears from C57Bl/6 mice were injected intradermally with 250 ng of IL-1 alpha , IL-17A, IL-22, OSM and TNF alpha (M5) or with Phosphate Buffered Saline (PBS). At 24 h quantitative RT-PCR analysis was carried out on total RNA and expression levels for cytokeratin 1 (CK1), cytokeratin 10 (CK10), loricrin (LOR), filaggrin (FLG), desmoglein 1 (DSG1), desmocollin 1 (DSC1) and involucrin (IVL) were normalized using GAPDH housekeeping gene and expressed as the fold decrease under non injected skin. Data are represented as mean and SEM of 3 independent experiments. *p<0.05, **p<0.01, ***p<0.001. (B) On day 2, the ears were collected for staining with Hematoxylin and Eosin (HE) and immunodetection of cytokeratin 10, loricrin, filaggrin, cytokeratin 6 and Ki-67. Scale bar 100 µm. Results are from one experiment representative of three. (C) Skin biopsies from normal control skin (Cont) or lesional psoriatic skin (Pso) were collected. Skin sections were stained with Hematoxylin and immunodetection of cytokeratin 10, loricrin, filaggrin, involucrin and S100A7 was performed. Scale bar 100 µm. Results are from one experiment representative of three. Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.pone.0101937), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

MicroRNA-214-3p (miR-214) mimic in rat vascular smooth muscle cells (A7r5) suppressed angiogenesis and proliferation but promoted senescence. a Schematic of the protocol for transfection of A7r5 VSMCs with a miRNA mimic control (mimic NC) or a miR-214 mimic and processed at the indicated times. b The effects of miR-214 mimic on VSMC cell proliferation were tested by CCK-8 assay. c Angiogenetic mRNA expressions of NOS3, VEGFA, CXCL12 and CXCR4 compared in cells transfected with mimic NC or miR-214 mimic (n = 3). d Senescence-associated mRNA expressions of TERT, TERF1 and TERF2 compared in cells transfected with mimic NC or miR-214 mimic (n = 3). e,g Representative western blots depicting TERF1, TERF2, p16INK4, p21CIP1, pRB, and quaking expression in mimic NC or miR-214 mimic transfected cells. f,h Normalized expressions of TERF1, TERF2, p16INK4, p21CIP1, pRB, and quaking (n = 3). i Senescence-associated beta -galactosidase staining demonstrating senescence in mimic NC or miR-214 mimic transfected cells. j Bar graphs show quantification of relative of SA-beta -gal positive cells (n = 3). Data are presented as the means ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001 (Two-tailed Student’s t-test) Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32410577), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Intraperitoneally Administered Compound A Effectively Suppresses Triple-Negative Breast and Blood Cancer Xenograft Tumor Growth. Immunocompromised mice were subcutaneously injected with cancerous cell lines and tumors were allowed to establish. Treatments occurred every other day and the studied compound or the equivalent vehicle control administered intraperitoneally for five weeks. Tumor volume and mass were measured two times per week. (a) MV-4-11 xenograft study was administered with approximately 5.0 mg/kg of Compound A and 5.0 mg/kg of curcumin. Results are representative for the Control (n = 5), Compound A (n = 5), and curcumin (n = 4) treated group, each with two tumors per mouse. (b) MDA-MB-231 xenograft study was administered with approximately 7.5 mg/kg of Compound A. Results are representative for the Control (n = 3) and Compound A (n = 4), each with one tumor per mouse. Scale bar is 1 centimeter. (c) Immunohistochemistry analysis of sectioned tumor tissues from the MDA-MB-231 study. Each section was subjected to the specified antibody followed by a biotinylated secondary antibody. Detection was done using a DAB Peroxidase HRP Substrate Kit (brown) followed by Hematoxylin counterstaining (purple). Images were obtained using inverted bright field microscopy. Sectioning results are representative of three individual tumors. Scale bar is 50 microns. Statistical analysis using One-Way ANOVA. *p < 0.05 vs tumor volume of the control. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28439094), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Mechanical tensile strain of 30% induces an elongation of paranodal junction and alteration in MAPK signaling in organotypic cerebellar slice culture. a Double immunostaining of organotypic cerebellar slices for CASPR in red and MAG in green. The response was evaluated at time 0 h post-stretch. Altered paranodal junctions are highlighted within white circles in high-magnification images. Scale bar, 50 μm; inset scale bar, 5 μm. b Sketch of CASPR and MAG labeling in the paranodal junction. Quantitative analysis is characterized by three length measurements: c nodal length called (A); d paranodal length called (B); and e (B-A) corresponding to the total length of the CASPR labeling. f–h Proteins were extracted following strain of 30% from control (Ctrl) and stretched (Stretch) slices and western blot was performed for the myelin protein MBP (f), ERK1/2 (g), and p38 (h). The ratio P-ERK on total ERK and the ratio P-p38 on total p38 are presented. beta -Actin was used as a loading control. Results represent the mean ± SEM (n = 3). ***p < 0.001. CASPR contactin-associated protein, MAG myelin-associated glycoprotein Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30298339), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Gomisin N (GN) represses the expression of JMJD2B, PPAR gamma 2, and PPAR gamma 2 steatosis target genes in HepG2 cells. HepG2 cells were treated with GN (100 μM) for 12 h. (A) JMJD2B expression was assessed by qPCR and western blotting. The full-length western blots corresponding to truncated blots are given in Supplementary Figure S4. (B) PPAR gamma 2 expression was measured by qPCR. (C,D) The expression of PPAR gamma 2 steatosis target genes was assessed by qPCR. Data represent means ± SEM of three independent experiments performed in triplicate. *p < 0.05 vs. no treatment. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30214048), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

TCAB1 knockdown inhibits tumor growth in vivo. a. shTCAB1-C and shScra cells were injected subcutaneously into the left and right neck of the nude mice respectively. Measured the tumors size from the 11th day using vernier caliper and calculated the volume of tumors by formula l*w*h/2 (mm3). Finally, got out the xenografts carefully after mercifully killed the mice. b. Measured the final volume and weighted the final weight of the removed tumors. c. Performed IHC against TCAB1 using mice xenografts sections. The smaller tumors expressed less TCAB1 compared to shScra cells. Statistical analysis of the IHC data used Aperio ImageScope software and all of the data were determined by Student’s t test (*P < 0.05, **P < 0.01, ***P < 0.005). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/25070141), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

MiR-375-3p negatively regulates Derlin-1 and blocks EMT in BFTC909 cells. (A) Western blot revealed the restoration of Derlin-1, MMP-2, Snail, and ZEB1 after co-transfection of miR-375-3p mimics and CMV-Derlin-1 compared with cells transfected with miR-375-3p alone in BFTC909 cells with alpha -tubulin as a reference (B) Quantification of the protein levels of Derlin-1, occludin, MMP-2, Snail, and ZEB1 from (A) (N = 3). (C) miR-375-3p suppressed BFTC909 cell migration ability but restored by Derlin-1 overexpression (N = 3). (D) miR-375-3p repressed invasion of BFTC909 cells but restored by Derlin-1 overexpression (N = 3). Data were represented as mean ± SD; * p < 0.05, ** p < 0.01. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35205628), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

TLR4 expression in pVW-MSCs cultured with or without LPS (10 μg/ml) for 1 and 4 h and after additional 24 h of recovery after LPS removal (4hR). a: flow cytometry analysis were performed in not fixed and not permeabilized cells for TLR4 surface expression determination (TLR4: Surface) and in fixed and permeabilized cells to measure the overall TLR4 amount (TLR4: Total). Red histograms: stained cells; blue histograms: control cells. b: representative Western Blot of TLR4 and housekeeping beta -tubulin and relative quantification were presented. c: representative images of TLR4 immunostaining of pVW-MSCs cultured with or without LPS (10 μg/ml) for 4 h. pVW-MSCs nuclei were stained with Hoechst 33258 (blue). Scale bar = 10 μm. Data shown represent the mean ± SD of three biological replicates, each experiment is repeated three times. Data were analysed using one-way ANOVA followed by the Tukey’s post hoc comparison test. Different letters above the bars indicate significant differences (p < 0.05). (AU = Arbitrary Units) Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31029157), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Ca, Ch, ChHa, Eq, Ha, Md, Pm, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ca, Ch, ChHa, Eq, Ha, Md, Pm, Rb

Applications:

, , , , , , , , , , , , , , , ,

Applications: , , , , , , , , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB400-105AF350
Alexa Fluor 405	NB400-105AF405
Alexa Fluor 488	NB400-105AF488
Alexa Fluor 532	NB400-105AF532
Alexa Fluor 594	NB400-105AF594
Alexa Fluor 647	NB400-105AF647
Alexa Fluor 700	NB400-105AF700
Alexa Fluor 750	NB400-105AF750
Allophycocyanin	NB400-105APC
Allophycocyanin/Cy7	NB400-105APCCY7
Biotin	NB400-105B
DyLight 350	NB400-105UV
DyLight 405	NB400-105V
DyLight 488	NB400-105G
DyLight 550	NB400-105R
DyLight 594	NB400-105DL594
DyLight 650	NB400-105C
DyLight 680	NB400-105FR
DyLight 755	NB400-105IR
FITC	NB400-105F
HRP	NB400-105H
Janelia Fluor 549	NB400-105JF549
Janelia Fluor 646	NB400-105JF646
PE	NB400-105PE
PE/Atto594	NB400-105PEATT594
PE/Cy5.5	NB400-105PECY55
PE/Cy7	NB400-105PECY7
PerCP	NB400-105PCP
mFluor Violet 450 SE	NB400-105MFV450
mFluor Violet 500 SE	NB400-105MFV500
mFluor Violet 610 SE	NB400-105MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB400-105SS

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NB600-1384

LC3B Antibody - BSA Free

(19 Reviews)

(397 Publications)

0.1 ml

NB600-1384

FTC-133/TPC-1 cells were transfected with HMGB1 shRNA and control shRNA and then pre-treated for 1 h with pepstatin A (PA, 10 uM) and E64D (10 uM) as indicated. Cells were subsequently treated for 3 h with HBSS in continuous presence or absence PA/E64D inhibitors. LC3-I/II levels were assayed by Western blot. Image collected and cropped by CiteAb from the following publication (//jeccr.biomedcentral.com/articles/10.1186/s13046-019-1328-3) licensed under a CC-BY license.

LC3B immunohistochemical staining in CC tissue(A) CC with low LC3B dot like staining. (B) CC with high LC3B dot like staining. (magnification 20X). Image collected and cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.17554) licensed under a CC-BY license.

Analysis of LC3B in treated U87-MG (human glioblastoma astrocytoma) lysates using anti- [Catalog # NB600-1384].

Depletion of RARalpha prevents ATRA-induced autophagic flux in SKBR3 cells. LC3B western blotting and quantification of the autophagic activity in control and RARalpha-knockdown SKBR3 cells upon treatment with 1 uM ATRA for 2 days in the presence or absence of BafA for 2 h at 200 nM. LC3B-II expression was normalized to GAPDH and to vehicle-treated control cells. Standard deviations from at least five independent experiments are shown. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2015236), licensed under a CC-BY license.

Human cervical carcinoma (HeLa) and Mouse Neuroblast cells (Neuro2a) were treated with (+) and without (-) 50 uM Chloroquine overnight. Whole cell protein lysates were prepared in 1x Laemmli sample buffer and approximately 10 ug of each lysate (NBP2-49689 and NBP2-49688) was separated on a 4- 20% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% nonfat milk in TBST. The membrane was probed with 2 ug/mL anti-LC3 (NB600-1384) and 1 ug/mL anti-alpha tubulin (NB100-690) as a loading control, and detected with the appropriate secondary antibodies using NovaLume Pico Chemiluminescence Substrate (NBP2-61915).

<b>Biological Strategies Validation. </b>Fibroblasts were exposed to 1 mM LLOMe and when indicated pretreated with the autophagy inhibitors 3-methyladenine (5 mM, 1 h) or bafilomycin A1 (25 nM, 30 min). Immunostaining with corresponding colocalization analysis of a) LC3 (green) and galectin-3 (red) and b) LC3 (green) and LAMP2 (red). Merged images show colocalization in yellow, nuclei are stained with DAPI (blue). LLOME = l-leucyl-l-leucine methyl ester, a lysosomal membrane damage compound. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32409651/) licensed under a CC-BY license.

Confocal analysis of HeLa cells using Rabbit anti-LC3B antibody (Catalog # NB600-1384, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).

Immunocytochemical/Immunofluorescent staining of treated U373-MG cells using the HRP conjugate of anti- (Catalog # NB600-1384). The nuclei were stained with DAPI.

Staining of treated U373-MG (human glioblastoma) cells using anti- [Catalog # NB600-1384].

Analysis of U87MG glioma xenografts using anti- [Catalog # NB600-1384]. Image from verified customer review.

LC3B staining in treated U87-MG cultured & subcutaneous tumors.

LC3B staining in glioblastoma multiform tissue.

Lane view shows a specific band for LC3B in 0.5 mg/ml of Neuro2A lysate at a molecular weight of approximately 15 kDa. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

LC3A and LC3B siRNAs specifically block the expression of the LC3A and LC3B proteins, respectively, in A549 cell line (E1,2,3). In E4 the reactivity of and of the LC3B (5F10 antibody) is shown, following silencing of the LC3A or of the LC3B genes, in the A549 cell line. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0137675), licensed under a CC-BY license.

NOD1-dependent induction of autophagy contributes to IRGM and LC3 recruitment to Mtb-containing vesicles. The subcellular localization of autophagy proteins was observed in untreated (A, C) and Tri-DAP treated cells (B, D) using anti-IRGM and anti-LC3 antibodies, which were detected using a secondary antibody coupled to 5 nm gold particles (indicated with arrowheads; TEM X 62,000). Gold particles colocalizing with bacteria were manually counted in 10 macrophages of each condition (E, F) and the differences between the treatments are indicated: ***p < 0.01 and *p < 0.05 vs. Mtb, ++p < 0.01 vs. Tri-DAP, Wilcoxon Rank test. Box plots indicate median and quartiles. Image collected and cropped by CiteAb from the following publication (//bmcpulmmed.biomedcentral.com/articles/10.1186/1471-2466-14-152), licensed under a CC-BY license.

Examples of immunohistochemical stainings; (A) LC3B low, (B) LC3B high, (C) p62 low (any cellular compartment), (D) p62 cytoplasmic low, (E) p62 cytoplasmic/dot like high, nuclear low, (F) p62 cytoplasmic/dot like low, nuclear high, (G) HMGB1 low, (H) HMGB1 high. Objective magnification, 40×.

Validation of modulations in autophagy-related markers after SIRT6 knockdown in melanoma cells. (B) Western blot analyses of various autophagy markers including LC3 I and II, BECN1, SQSTM1, ATG3, ATG7, ATG10 and GAA using cell lysates of shNS and shSIRT6 in A375 melanoma cells. beta-actin was used as loading control. The figure shows representative western blot images from 2-3 experiments. Image collected and cropped by CiteAb from the following publication (//www.genesandcancer.com/lookup/doi/10.18632/genesandcancer.153), licensed under a CC-BY licence.

Induction of autophagic flux in OE19 upon Lapatinib treatment. (a) LC3B flux was assessed comparing control and BafilomycinA (BafA)-treated (200 nM, 2 h) OE19 upon Lapatinib treatment (120 nM, 24 h). LC3 band intensities were quantified using ImageJ software. Total protein was used as a loading control, and Phospho-Her2 for Lapatinib treatment (n = 3). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30297650), licensed under a CC-BY licence.

In vitro alphaS seeds induce endo-lysosome-free and endo-lysosome-associated alphaS inclusions in H4/V1S-SV2 cells. (d) The treated cells were further subjected to immunocytochemical staining of LAMP1 (Alexa fluor 405) and LC3 (Alexa fluor 568) to demonstrate the association between autophagosomes, endo-lysosomes, and seeded alphaS inclusions. Scale bar: 5 um (top row) and 2 um (bottom row). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-08149-w), licensed under a CC-BY licence.

(E) LC3A and LC3B siRNAs specifically block the expression of the LC3A and LC3B proteins, respectively, in A549 cell line (E1,2,3). In E4 the reactivity of LC3A (ab62720 Ab) and of the LC3B (5F10 antibody) is shown, following silencing of the LC3A or of the LC3B genes, in the A549 cell line. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26378792), licensed under a CC-BY licence.

Combination treatment of valproic acid (VPA) and doxorubicin (DOX) synergistically augmented the autophagy of HepG2 cells. (D) LC3 I and II protein levels were analyzed using Western blotting. Actin was used as the loading control. The intensity of LC3B-II bands was quantified by scanning densitometry program ImageJ and normalized to that of actin (right panel). Three independent experiments were performed and results reported as the mean +/- standard deviation (SD). ** p < 0.01, *** p < 0.001 compared with the control group. Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/1422-0067/18/5/1048), licensed under a CC-BY licence.

Autophagy and the proteasome regulate CagA stability. (A) Wild-type (WT) and autophagy-deficient (Atg5-/-) MEFs were infected with a CagA+ vacA- isogenic mutant strain (MOI 100) for 8 hours using a gentamycin protection assay. The autophagy marker LC3-II was used to confirm that Atg5-/- MEFs are autophagy-deficient. CagA protein levels were measured by Western blotting using beta-actin as loading control. Graph shows fold change of CagA normalized to beta-actin relative to WT MEFs (mean + SEM; n = 4). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30631092), licensed under a CC-BY licence.

PB induces autophagy but blocks autophagosome trafficking and autophagosome-lysosome fusion in Hep3B cells. (B) Hep3B cells were treated for 6, 24 and 48 h with the indicated concentrations of PB. Levels of protein expression were analysed by Western blot using antibodies against LC3B, p62/SQSTM1, ATG12, BECN1 and Actin. Images were cropped from different blots run under the same experimental conditions. The original blots were attached as Supplementary Figure 3A. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep41437), licensed under a CC-BY licence.

Effects of exogenous HMGB1 on ATRA-induced differentiation. E). Expression of PML-RARalpha, P62 and LC3-I/II in NB4 cells treated with ATRA (1 uM) or HMGB1 (10 ug/ml) for 48 h was assayed by western blotting and co-immunoprecipitation in comparison to control (DMSO). Image collected and cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.15432), licensed under a CC-BY licence.

Activation of autophagy by MG132 is associated with AGR2 decline. d) The levels of AGR2 and LC3B were measured by western blotting with MG132 treatment at indicated time. e Analysis of LC3B and AGR2 accumulation in A549 cells by fluorescent staining. Scale bar = 10 um Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30647455), licensed under a CC-BY licence.

Autophagy was induced under conditions of LH and inhibited by CQ and 3-MA in HCC cells.(A-C). SMMC-7721 and HepG2 cells were directly incubated under normal or LH conditions for 8 h or incubated under LH conditions in the presence or absence of CQ or 3-MA for 8 h. (C). The whole-cell lysates were subjected to western blot analysis as described in the 'Materials and methods'section. The experiments were repeated at least three times. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep05382), licensed under a CC-BY licence.

The effect of H19 siRNA on autophagy in OGD/R model. A) Immunofluorescence of LC3II in different conditions. Image collected and cropped by CiteAb from the following publication (//www.aginganddisease.org/EN/10.14336/AD.2016.0530), licensed under a CC-BY licence.

Validation of selected differentially expressed genes. A. RT-PCR analysis of genes elevated in drug resistant cells. The y-axis represents fold up-regulation in the different drug resistant cell lines over the parental OV90 cell line. B. RT-PCR analysis of genes decreased in drug resistant cells. The y-axis represents the fold down-regulation of the different resistant cell lines compared to the parental OV90 cell line. C. Immunoblot analysis of selected gene products identified by microarray and RT-PCR as altered in drug resistant cells. Image collected and cropped by CiteAb from the following open publication (//ovarianresearch.biomedcentral.com/articles/10.1186/1757-2215-4-21), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Osteogenic differentiation in 3D. (A) Histochemical staining of 12-μm-thick gel sections from one representative donor after 5 weeks of culture in undifferentiated or osteogenic conditions. Osteoimage® (HA-specific staining), von Kossa (stains anionic portion of phosphates, carbonates, and other salts) and Alizarin Red (stains Ca2+ deposits) shows differentiation in the Col I—BMSC cultures. Scale bar = 100 μm. (B) Immunostaining for osteocalcin (OCN) and nuclear 4′,6-diamidino-2-phenylindole (DAPI) in 3D cultures after 5 weeks. (C) Immunostaining for osteopontin (OPN) and nuclear DAPI in 3D cultures after 5 weeks. Scale bar = 50 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34948393), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

G-TPP induces PINK1 stabilization and kinase activity in HeLa cells(A) G-TPP treatment leads to PINK1 stabilization and pS65-Ub induction in HeLa cells. HeLa cells stably expressing untagged Parkin were treated with 10 µM G-TPP for the indicated times. Western blots were prepared with cell lysates and probed with antibodies against PINK1 and pS65-Ub. GAPDH served as a loading control. (B) pS65-Ub is induced in G-TPP treated cells and co-localizes with EGFP-Parkin and mitochondria. HeLa cells stably expressing EGFP-Parkin (green) were treated with 10 µM G-TPP for the indicated times and fixed. Cells were stained with antibodies against pS65-Ub (red) and the mitochondrial marker TOM20 (cyan). Scale bars correspond to 10 µM. (C) Quantification of Parkin translocation using High Content Imaging. HeLa EGFP-Parkin cells were treated for 4 or 8 h with or without 10 µM G-TPP. CCCP treatment (10 µM for 2 h) was used as a positive control. Cells were fixed, counterstained with Hoechst dye to visualize nuclei, imaged and analyzed using the ratio of cytoplasmic to nuclear EGFP signal [21]. Data was normalized to positive (2 h 10 µM CCCP treatment) and negative (2 h DMSO) controls. G-TPP significantly induced Parkin re-localization to levels similar to or beyond 2 h CCCP treatment. Shown are the mean values of three independent experiments with triplicate wells each ± SEM (one-way ANOVA with Tukey’s posthoc, ***p < 0.0005). Image collected and cropped by CiteAb from the following open publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.22287), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Association of p53 and APE1 on p53-binding sites in p21 promoter.(A) p21 promoter structure showing p53 and AP4 binding sites. (B) ChIP Real Time PCR analysis showing relative enrichment (2− delta CT) of APE1-immunoprecipitated DNA over that from control IgG in p21 promoter regions containing p53 binding sites 1 & 2 in HCT116WT cells. (C) Re-ChIP analysis (first IP with alpha -APE1 and the second IP with alpha -p53 antibody) showing simultaneous recruitment of APE1 and p53 in control vs. etoposide treated cells; *: p value <0.05 (n = 2) calculated based on APE1/p53 enriched DNA from control vs. etoposide treated cells. (D) Western analysis of FLAG immunoprecipitate (IP) to detect APE1-associated p53 and FLAG (APE1) from empty vector vs. FLAG-tagged WT APE1 or FLAG-tagged N delta 33 APE1 transfected HCT116WT cells (left panel) and from control vs. etoposide-treated WT APE1-FLAG transfected cells (right panel). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/23874636), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Loss of functional PRPF31 induce RNA:DNA hybrid dependent genomic instability but not in mice retinal neurons.(a andb) gamma H2AX and 53BP1 foci analysis in PRPF31 siRNA-transfected RPE-1 cells. (c and d) gamma H2AX and 53BP1 foci analysis in vasculo-stromal fraction derived primary cells fromPrpf31+/A216P mice (Prpf31-ki). (e) gamma H2AX and 53BP1 foci analysis in retina fromPrpf31+/A216P mice on postnatal day 20. All column bars represent the mean. For (a-d) “n”, mentioned on respective column, signify number of cells analyzed from two independent experiments. For (e) n=16 for each column and signify number of retinal sections analyzed; acquired from n=4 eyes. Error bars represent Standard error of Mean (SEM). *P≤0.05; **P<0.01, ***P<0.001 using Mann-Whitney test (a,b), Kruskal-Wallis test followed by Dunn’s post hoc test (c,d); and two tailed unpaired Student’s t-test (e). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30345028), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Subarachnoid hemorrhage (SAH) grades and temporal expression of endogenous osteopontin (OPN) and autophagy‐related proteins in rat brain after SAH. A, Representative brain images of Sham and SAH rats. B, Summary of SAH grading scores of all groups. Sample size is 27, n = 9 per group. Data were analyzed using Kruskal‐Wallis test, χ2 = 18.183. C, Representative Western blot images and quantitative analyses of OPN, Beclin 1, LC3, and ATG 5 from the left hemisphere of rat brains at different time points after SAH. Sample size is 36, n = 6 per group. Data were presented as mean ± SD. F = 28.45 for OPN, F = 12.37 for Beclin 1, F = 18.88 for LC3, F = 22.14 for ATG5. *P < .05, ** P < .01, ***P < .001 vs Sham group. SAH, subarachnoid hemorrhage; Vehicle, phosphate‐buffered saline; rOPN, recombinant OPN. NS, not significant Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31436915), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Staufen1 co-fractionates with Gag and vRNA in gradient density fractionation analyses. (A) HeLa cells were either mock transfected with empty vector, pcDNA3, or with a plasmid that expresses Staufen1-HA. The transfected cells were collected 24 h later, lysed and were either mock-treated or treated with RNAse A. The lysates were then fractionated on 5–50% sucrose gradients and 20 fractions were collected for further analysis by western blotting for viral and host proteins, as indicated. HIV-1 viral genomic RNA (vRNA, 9 kb) was assessed in each fraction by slot blot analysis. TL represents the total lysates. (B) HeLa cells were co-transfected with pNL 4–3 and Staufen1-HA. The presence of Staufen1-HA, precursor Gag and p24 were assessed in each fraction by western blotting analysis. HIV-1 viral genomic RNA (9 kb, vRNA) was assessed in each fraction by slot blot analysis. Staufen1, Gag and vRNA were quantitated in each fraction by densitometry and relative levels are depicted for each fraction (Blue: Staufen1-HA, Red: Gag, Green-vRNA). Image collected and cropped by CiteAb from the following open publication (//journal.frontiersin.org/article/10.3389/fmicb.2012.00367/abstract), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Endogenous MIR376A limits starvation-induced autophagy.(A) Blockage of endogenous MIR376A by Ant-376a, but not CNT-Ant further stimulated starvation (STV)-activated LC3-I to LC3-II conversion in MCF-7 cells. ACTB was used as a loading control. LC3-II/LC3-I densitometric ratios were marked. (B) Ant-376a, but not CNT-Ant resulted in further activation of SQSTM1 protein degradation following starvation in MCF-7 cells. SQSTM1/ACTB ratios were marked. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/24358205), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

GAS6 increases the amount of both CIP2A and p-DAPK in RSC96 cells(A) RSC96 cells were incubated with GAS6 (100 ng/ml), and immunoblotting evaluations of CIP2A and p-DAPK expression were performed. The lower panel shows the intensity during basal conditions from the scanned imaged (upper panel). (B) RSC96 cells incubated with and without GAS6 (100 ng/ml) for 30 min and analysed by immunofluorescence. (C) Immunohistochemistry images demonstrated that CIP2A expression levels were higher than sham levels at 14 days and 28 days in the cavernous nerve after BCNI. Image collected and cropped by CiteAb from the following open publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.23978), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Effect of treatment with FX extract on CREB/BDNF signaling. (a-c) Differences in the phosphorylation of CREB and expression of BDNF between groups in the PFC and hippocampus (n = 3-4). (d, e) pCREB and BDNF immunofluorescence was assessed in the PFC and hippocampus (n = 3-4). Mean ± SD. #P < 0.05 versus normal group; ∗P < 0.05 and ∗∗P < 0.01 versus control group. Scale bar = 200 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30065945), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry of seven in absentia homolog 2 (SIAH2) gene in normal, in situ and invasive breast carcinomas. (A) Occasional nuclear positivity (arrows) in luminal cells in the terminal duct lobular unit. (B) Moderate to strong staining of SIAH2 in the nucleus of a small proportion of the cell in a high nuclear grade ductal carcinoma in situ with comedo necrosis. (C) Occasional weak to moderate SIAH2 (arrows) staining in a luminal type ductal carcinoma. (D) Strong SIAH2 staining in all nuclei in this basal-like breast carcinoma. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/21306611), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

CNF1-triggered IL-1 beta maturation requires activated Rac, ASC and caspase-1.(A) Western blot analysis of the production and maturation/secretion of IL-1 beta by primary macrophages following treatment with CNF1, LPS or CNF1+LPS for 10 h. Actin and BSA were used as loading controls. (B) Quantification of caspase-1 activity in macrophages following treatment with CNF1+LPS for 6 h using YVAD-Fluorescent Labelled Inhibitor Caspase-1 Activity (FLICA). (C) Western blot analysis of macrophages IL-1 beta maturation/secretion upon transfection of HA-Rac2Q61E and LPS treatment. (D) Co-immunoprecipitation of Myc-Rac2 and caspase-1 using an anti-Myc antibody following the treatment of HEK 293T cells with CNF1+LPS for 6 h. Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.ppat.1004732), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Expression of sphingosine-1-phosphate (S1P), interleukin (IL)-1 beta , and protease-activated receptor-1 (PAR-1) in isolated astrocytes. (A) Isolated and purified astrocytes were identified by bright-field microscopy and immunofluorescence for glial fibrillary acid protein (GFAP; red). Nuclei (DAPI) were labeled in blue. Scale bar = 100 μm. (B) Western blot and quantitative reverse transcription polymerase chain reaction (qRT-PCR) of the protein and mRNA expression, respectively, of S1P 0, 1, and 6 h after astrocytes were treated with lipopolysaccharide (LPS) or thrombin and Dabigatran (Dab) or PAR-1-inh (LPS only). (D) qRT-PCR and (E) enzyme-linked immunosorbent assay (ELISA) of the mRNA expression and secretion, respectively, of IL-1 beta 0, 1, and 6 h after astrocytes were treated with LPS or thrombin and Dab or PAR-1-inh (LPS only). (F) Immunofluorescence of astrocytes and quantification of relative mean integrated optical density (IOD; green fluorescence). PAR-1 was stained in green and nuclei (DAPI) were stained in blue. Scale bar = 100 μm. For (B–F), data are presented as the mean ± SD (n = 3). *p < 0.05; #p < 0.05 vs. the same group at 0 h; &p < 0.05 vs. the same group at 1 h. ns, not significant. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32694981), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

53BP1/RIF1 work epistatically to counteract BRCA1/CtIP-mediated end resection to regulate DSB repair pathway choiceA. HeLa cells harboring pGC were treated with the indicated siRNAs before transfection with I-SceI-expressing vector to induce DSBs. After 48h, the percentage of GFP+ cells was measured as an indication for HR efficiency. B.-D. Asynchronous A549 cells were treated with the indicated siRNAs before irradiation with 2Gy and CtIP B. and RPA foci C. were monitored at 2h while RAD51 D. foci were enumerated at 4h. E. Left panel: representative photos for the colocalization between 53BP1 and RIF1 foci in A549 cells at 2h after 2Gy. Right panel: quantitation of 53BP1, RIF1 or colocalized foci in the absence (DMSO) or presence of ATM inhibitor (ATMi). F. Left panel: representative micrographs for 53BP1 or RIF1 foci in irradiated cells after depletion of either proteins. Right panel: quantitation of experiments presented in the left panel. At least 100 nuclei were counted. In all cases, the number of foci measured in non-irradiated cells was subtracted (relative). Shown are the mean ±SEM for three independent experiments. sc: scrambled RNA, si53: si53BP1, siR: siRIF1, siB: siBRCA1, and siC: siCtIP. Image collected and cropped by CiteAb from the following open publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.11023), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Up-regulation of RAS induced by cholesterol and 25-hydroxycholesterol in HK-2 cells. HK-2 cells were made quiescent by serum-free medium for 24 hours and then maintained in serum-free medium (control) or serum-free medium containing 30 μg/ml cholesterol together with 1 μg/ml 25-hydroxycholesterol (lipid) for 24 hours. (A) Real-time PCR of the total RAS components mRNA prepared from HK-2 cells with or without lipid treatment. AT1/AT2 ratio was evaluated as the balance between AT1 and AT2. beta -actin was used as mRNA loading control. (B) Western blot analysis for RAS components protein expression. (C) The histogram shows the average volume density corrected by the housekeeping control, beta -actin. Data is expressed as mean ± SD. *P < 0.05 vs. control. Angiotensinogen, AGT; angiotensin II, Ang II; angiotensin converting enzyme, ACE; angiotensin II type 1 receptor, AT1; angiotensin II type 2 receptor, AT2. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/23570453), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Effect of TERT silencing on murine M1 macrophage polarization.TERT siRNA and GV144-TERT were transiently transfected into LPS-treated RAW264.7 cells, respectively. (a) The endogeous TERT levels were detected by real-time PCR and western blot. The results are shown as relative expression against control expression without treatment. (b) The mRNA levels of M1 macrophages biomarkers including TNF-alpha , IL-1 beta , NOS2 and CCL2 were detected by real-time PCR. The results are shown as relative expression against control expression without treatment. (c) The secretion of proinflammatory cytokines including TNF-alpha , IL-1 beta , IL-6 and IL-12 were determined by ELISA. (d) TERT successful over-expression was verified by real-time PCR and western blot in LPS-stimulated RAW 264.7 cells. The results are shown as relative expression against control expression without treatment. (e) The mRNA levels of M1 macrophages biomarkers were detected by real-time PCR. The results are shown as relative expression against control expression without treatment. (f) The secretion of proinflammatory cytokines were determined by ELISA. Data shown are the mean ± SD from 3 independent experiments. *P < 0.05, **P < 0.01 vs control group. #P < 0.05, ##P < 0.01 vs LPS-treated group. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/26725521), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Ang II increases the number of TSP-1 positive cells in mouse heart and skin.IHC staining of TSP-1 was performed on paraffin sections from the heart (A) and skin (B) of PBS and Ang II treated WT mice. Representative photographs are shown from five animals per group. C. TSP-1 protein levels were increased in Ang II treated HDMECs in a dose dependent manner (*p≤0.05). Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.pone.0109763), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Ba, Bv, Ca, Pm, Ye, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ba, Bv, Ca, Pm, Ye, Ze

Applications:

, , , , , , , , , ,

Applications: , , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB600-1384AF350
Alexa Fluor 405	NB600-1384AF405
Alexa Fluor 488	NB600-1384AF488
Alexa Fluor 532	NB600-1384AF532
Alexa Fluor 594	NB600-1384AF594
Alexa Fluor 647	NB600-1384AF647
Alexa Fluor 700	NB600-1384AF700
Alexa Fluor 750	NB600-1384AF750
Allophycocyanin	NB600-1384APC
Biotin	NB600-1384B
DyLight 350	NB600-1384UV
DyLight 405	NB600-1384V
DyLight 488	NB600-1384G
DyLight 550	NB600-1384R
DyLight 594	NB600-1384DL594
DyLight 650	NB600-1384C
DyLight 680	NB600-1384FR
DyLight 755	NB600-1384IR
FITC	NB600-1384F
HRP	NB600-1384H
Janelia Fluor 549	NB600-1384JF549
Janelia Fluor 646	NB600-1384JF646
PE	NB600-1384PE
PerCP	NB600-1384PCP
mFluor Violet 450 SE	NB600-1384MFV450
mFluor Violet 500 SE	NB600-1384MFV500
mFluor Violet 610 SE	NB600-1384MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB600-1384SS

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NB100-1028

AIF-1/Iba1 Antibody

(21 Reviews)

(395 Publications)

0.1 mg

NB100-1028

GM1 effects on brain microglia in vivo, after peripheral administration of LPS. Immunoblot in whole brain lysates and densitometric analysis show LPS-dependent increase in AIF-1/Iba1 expression. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/34991625/) licensed under a CC-BY license.

Reactive microglia (green) in peritumoral area of IC injected human glioblastoma cells in mouse brain. NeuN (red), DAPI (blue). Antibody at 1:250. IHC image submitted by a verified customer review.

AIF-1/Iba1 in canine brain. Primary antibody used at 1 ug/mL and incubated with tissue for 30 minutes at room temperature. IHC-P image submitted by a verified customer review.

Staining in mouse brain using AIF-1/Iba1 Antibody. IHC-P image submitted by a verified customer review.

Brown arrow= AIF-1/Iba1 positive cells, Green arrow= mTOR positive cells, Black arrow= mTOR and AIF-1/Iba1 positive cells. IHC-P image submitted by a verified customer review.

Staining in rat hippocampus using AIF-1/Iba1 Antibody. IHC-Fr image submitted by a verified customer review.

Feline optic nerve microglia were immunolabeled with AIF-1/Iba1 Antibody (green), DAPI (blue). Imaging was performed using an epifluorescence microscope. IHC-Fr image submitted by a verified customer review.

Mouse dorsal root ganglia. AIF-1/Iba1 expression in reactive macrophages. Maximum intensity z-projection from 3D images. IHC image submitted by a verified customer review.

Staining in mouse brain using AIF-1/Iba1 Antibody. IHC image submitted by a verified customer review.

Analysis of LPS injected mouse brain tissue, 20 um thickness, frozen. Primary antibody dilution, 1:200. Secondary antibody CY3. Mounted with DAPI mount. Image from verified customer review.

AIF-1/Iba1 staining of AD mouse model (hippocampus). Image from verified customer review.

Negative Control showing staining of paraffin embedded Human Spleen, with no primary antibody.

(0.1ug/ml) staining of Mouse Lymph node lysate (35ug protein in RIPA buffer). Detected by chemiluminescence.

(1ug/ml) staining of Human Frontal Cortex (A) and (0.1ug/ml) Mouse (B) and Rat (C) Brain lysate (35ug protein in RIPA buffer). Detected by chemiluminescence.

(8ug/ml) staining of paraffin embedded Human Spleen. Heat induced antigen retrieval with citrate buffer Ph 6, HRP-staining.

GM1 effects on brain microglia in vivo, after peripheral administration of LPS. C) Immunoblot in whole brain lysates and densitometric analysis show LPS-dependent increase in Iba 1 expression. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/34991625), licensed under a CC-BY licence.

CXCR2+neutrophils are not required for microglial activation. a Representative images of Iba1 immunohistochemistry staining in lumbar spinal cord ventral horns of naive control, control EAE, and Cxcr2 cKO EAE mice. Quantitative analysis of Iba1+ cell (b) soma area as shown in a. A total of 2338 Iba1+ cells were included in our analyses of soma size (cont naive, 222 Iba1 cells; control EAE, 1236 Iba1+ cells; Cxcr2 cKO EAE, 880 Iba1+ cells). Control naive, n = 3; control EAE, n = 3; Cxcr2 cKO EAE, n = 3. *p 1.5, FDR-corrected p1.5, FDR-corrected p<0.05) to identify GO terms, including inflammatory response (b). The volcano plot shows significant DEGs derived from the NanoString analysis of the brains of Plcg2 inactivation (PLCG2 inact, n=3) and WT (n=3) mice (1-month male) (c). Immunostaining of microglial markers with IBA1 (green) and P2RY12 (red) and quantification of percent area in brain sections of 1-month-old male WT and PLCG2 inact demonstrate unchanged microglial morphology and area coverage (d). GO gene ontology, BP biological process, DEGs differentially expressed genes, FDR false discovery rate Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35180881), licensed under a CC-BY licence.

Deficiency of HDAC3 in microglia suppresses the cGAS-STING-mediated autoimmune response. (K) Proteins extracted from the ischemic penumbra and contralateral cortex of HDAC3 cKO and WT mice subjected to tMCAO 6 h post-reperfusion were analyzed with antibodies against cGAS, HDAC3, Iba1, and GAPDH. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32863951), licensed under a CC-BY licence.

ADT induced SPINK1 upregulation associates with NE-phenotype in mice and NEPC patients.a Box plots depicting relative expression of SPINK1, SYP, CHGA, TUBB3, and VIM transcripts (read counts) in VCaP tumors implanted orthotopically in orchiectomized mice and subjected to vehicle (n = 4) or anti-androgens [enzalutamide (n = 4) or ARN-509 (n = 4)] treatment for 4 weeks (GSE95413). b Representative images of immunohistochemical staining for the same markers shown in a using VCaP xenograft tumors as described in a. Scale bar represents 100 μm. c Box plots depicting quantification of the immunohistochemical staining in VCaP xenografts for the markers shown in b. d Representative images for immunohistochemical staining of SPINK1, SYP, CHGA, and TUBB3 in 22RV1 xenograft tumors excised from orchiectomized mice treated with enzalutamide (20 mg/kg body weight) or vehicle control (n = 5 each). Intact group represents non-castrated control mice (n = 5). Scale bar represents 50 μm. e Box plots depicting quantification of the immunohistochemical staining in 22RV1 xenografts for the markers shown in d. f Representative images showing H&E staining (×200 magnification) and immunostaining (×200 magnification) for AR, synaptophysin, and SPINK1 in tumor specimens obtained from NEPC patients’, namely WCM12, WCM155 (an organoid), and WCM677. Scale bar represents 100 µm. Data are presented as box-and-whisker plots with median, where the box extends from 25th–75th percentile, and whiskers ranges from minimum and maximum values. For panels a, c, e one-way ANOVA, Dunnett’s multiple-comparisons test was applied. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31959826), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

EBNA1 promotes the upregulation of oxidative DNA damage repair pathways in EBV converted cell lines and EBV positive BLs. a Representative western blots illustrating the expression of MTH1, OGG1, and MUTYH in pairs of EBV-negative and -positive cell lines. GAPDH was used as loading control. b Densitometry quantification of the specific bands. The intensity of the specific band in EBV positive cells relative the EBV-negative parental is shown. Mean ± SE of four independent experiments. c Representative western blots illustrating the expression of EBNA1, MTH1, OGG1, and MUTYH in the Mutu cell lines. d Densitometry quantification of expression in the EBV positive cell lines relative to the EBV-negative Mutu-30. Mean ± SE of four independent experiments Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31511648), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Rab31 expression levels in stably transfected breast cancer cells. (A) pAb RT3-IgG specifically detects rab31 in cell lysates from stably transfected MDA-MB-231 and CAMA-1 breast cancer cells in Western blot analyses. Vector control, MDA-MB-231, or CAMA-1 cells were stably transfected with the empty vector pRcRSV; batch, parental MDA-MB-231 (low endogenous rab31 expression) or CAMA-1 (no endogenous rab31 expression) cells were stably transfected with the pRcRSV-based rab31 expression vector; high, medium or low, selected cell clones of batch-transfected MDA-MB-231 or CAMA-1 cells with differing high, medium or low protein levels of rab31. (B) Rab31 antigen values (ng/mg of total protein) in rab31-transfected MDA-MB-231 and CAMA-1 cells, determined by rab31-ELISA. Mean values (± SEM) of at least three independent experiments are depicted. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/22920728), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

EGFR inhibition downregulates VEGF and HIF-1 in vitro and in vivo.SQ20B cells were seeded at 25% confluence and cultured overnight. The next morning, cells were transfected with 150 nanomoles siRNA (either EGFR SmartPool or control non-targeted) in Optimem transfection medium using Oligofectamine. After 24 hours, the transfection medium was replaced with fresh, prewarmed culture medium. 24 hours after this (48 hours after transfection), cells were exposed to either 21% or 1% oxygen. Three hours later they were harvested and Western blotting was performed. (B) SQ20B cells were treated with 10 nM cetuximab or DMSO (control) for 16 hrs, and then exposed to either 21% or 1% oxygen. Three hours later they were harvested and Western blotting was performed. (C) Nude mice were injected subcutaneously in the flank with SQ20B cells to form xenografts. When the tumors reached a size of 100–150 mm3 (approximately 7–10 days after injection), half the mice (lanes 4–6) were started on an erlotinib-containing diet (50 mg/kg/day). After 4 days of feeding, mice were sacrificed, and the tumors were removed. Half of each tumor was lysed in protein lysis buffer, and Western blotting was performed for pAkt (S473) and HIF-1 alpha . Each lane represents a tumor from a different mouse. (D) The other half of each tumor from (E) was homogenized in PBS, and then ELISA for VEGF was performed. VEGF level was normalized to tumor weights. Data shown represent mean values from 3 control mice and 3 erlotinib-treated mice. Error bars represent standard error of the mean. p value was obtained using Student's t-test. (E) H226 cells were treated with 10 µM erlotinib or DMSO (control) for 16 hrs, and then exposed to either 21% or 1% oxygen. Three hours later they were harvested and Western blotting was performed. (F) H226 cells were seeded then later that day they were treated with erlotinib (10 µM) or DMSO (control). 24 hours later, the media was changed and replaced with media containing 1% serum with or without erlotinib. Cells were exposed to 1% oxygen. 16 hours later, aliquots of supernatant were removed from dishes, and ELISA for VEGF was performed. ELISA values were normalized to the number of cells present. Data shown represent mean values. Error bars represent standard error of the mean. p value was obtained using Student's t-test. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/19657384), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Elevated levels of additional proteins and GM3 ganglioside in the MEC of MPS IIIB brain.Staining performed with antibodies to the indicated substances was observed in the MEC region of 3 month-old MPS IIIB mice (for total ubiquitin and polyubiquitin) and 6 months for all others. Staining was not seen in the MEC region of age-matched control mice (Naglu +/−) nor in the LEC region of MPS IIIB mice (the latter not shown). Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.pone.0027461), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

ERK2 utilizes kinase-docking domains to interact with ATG5–ATG12 and LC3-II.(a) Mutations in FRS on ERK2 decrease colocalization of ERK2 with ATG5–ATG12, LC3 and WIPI1. Immunofluorescence (IF) showing colocalization (depicted as white pixels by ‘colocalization finder application’) of WT-ERK2-HA, FRS ERK2 mutants (L198A-, L232A-, L198A/L232A-, Y261A-ERK2-HA) or common docking (CD) mutant (D319N-ERK2-HA) with ATG5–ATG12 (panels 1–6), LC3 (panels 7–12) or WIPI1 (panels 13–18) in EGF-treated NIH/3T3 cells. ERK2 is stained in red, and autophagy proteins are stained in green. Scale bar, 10 μm. The bars represent mean±s.e.m. **P<0.01, ****P<0.0001 compared with WT-ERK2-transfected cells; Student’s t-test, 50 cells analysed from n=2. (b) Mutations in FRS on ERK2 decrease colocalization of ERK2 with nuclear ATG5–ATG12 or LC3. IF showing colocalization (white pixels) of WT-ERK2-HA (red), FRS ERK2 mutants (L198A-, L232A-, L198A/L232A-, Y261A-ERK2-HA) or common docking (CD) mutant (D319N-ERK2-HA) with ATG5–ATG12 (panels 1–6) or LC3-II (panels 7–12) in EGF-treated NIH/3T3 cells. Scale bar, 5 μm. The bars represent mean±s.e.m. **P<0.01, ***P<0.001, ****P<0.0001 compared with WT-ERK2-transfected cells; Student’s t-test, 50 cells analysed from n=2. Arrows depict colocalization. (c) ATG5–ATG12 conjugation is required for ERK phosphorylation. Immunoblots for P-ERK, total ERK and beta -actin in NIH/3T3 cells transfected with WT ATG5 or the conjugation-defective ATG5 K130R mutant and treated with EGF (10 min). The bars represent mean±s.e.m. *P<0.05 compared with WT ATG5-transfected cells; Student’s t-test, n=3. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/ncomms3799), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Human tumor microarrays (TMAs).Pilocytic astrocytoma TSPO immunohistochemistry (A) and TSPO immunohistochemical scoring (C). Glioblastoma multiforme TSPO immunohistochemistry (B) and TSPO immunohistochemical scoring (D). Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.pone.0141659), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

DNMT1 reveals a SCMC-like cortical localization with aberrant localization in Nlrp2tm1a/tm1a derived oocytes.(A) Whole mount immunofluorescence co-staining with anti-DNMT1 and DAPI on the same Nlrp2M+/Z+ embryos used to stain NLRP2 reveals a SCMC-like cortical localization of DNMT1 with a diffuse nuclear stain in later stage embryos. Scale bar represented is 200 μm. (B) DNMT1 staining in paraffin embedded oocyte sections reveals a less cortical, more cytoplasmic intense stain with nucleolar focus in oocytes from Nlrp2tm1a/tm1a dams compared to oocytes from Nlrp2+/+ dams. (C) Whole mount immunofluorescence for DNMT3A in unfertilized control oocytes reveals a characteristic metaphase associated localization and no difference is noted in oocytes derived from Nlrp2tm1a/tm1adams. (D) Whole mount immunofluorescence in unfertilized oocytes reveals that as expected, DNMT3B is not yet expressed in unfertilized control oocytes and no expression is noted in oocytes derived from Nlrp2tm1a/tm1adams. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/srep44667), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Activation of MAPK7 in chondrocytes is essential for hypoxic adaptation and enhancement of HIF1 alpha signaling under hypoxia. a, b Immunofluorescence staining of (a) MAPK7 (b) P-MAPK7 on representative sections of distal femoral growth plates at P1. Scale bar, 200 μm. RZ, resting zone; PZ, proliferative zone; HZ, hypertrophic zone. c Western blot analysis of MAPK7 and p- MAPK7 in primary chondrocytes cultured under hypoxic conditions for 0, 1, 4, 12, or 24 h. d Immunofluorescence staining of SOX9 on representative sections of distal femoral growth plates at P1. Scale bar, 200 μm. e Western blot analysis of SOX9, COL2A1, MAPK7, HIF1 alpha , and VEGFA in CON and Mapk7 CKO chondrocytes cultured under normoxic or hypoxic conditions for 48 h. f Western blot analysis of SOX9, COL2A1, MAPK7, p-MAPK7, HIF1 alpha , and VEGFA protein levels in XMD8-92- or DMSO-treated primary chondrocytes cultured under normoxic or hypoxic conditions. Primary chondrocytes harvested from wild-type mouse knees were cultured in the presence of 5 μM XMD8-92 or an equal amount of DMSO under normoxia for 3 h, followed by normoxia or hypoxia for 48 h. XMD, XMD8-92. g, h mRNA levels of (g) Sox9 and (h) Col2a1 in CON and Mapk7 CKO chondrocytes cultured under normoxia or hypoxia were measured by real-time quantitative PCR (n = 3). *P < 0.05 (one-way ANOVA followed by Dunnett’s post hoc test). Data are presented as mean ± SD. i Immunofluorescence staining of HIF1 alpha on representative proximal tibial growth plate sections from P1 mice. Scale bar, 200 μm. j mRNA levels of Hif1a and its known target genes were measured in growth plate cartilage from P1 Mapk7 CKO mice by real-time quantitative PCR. k HIF1 alpha activity in CON and Mapk7 CKO chondrocytes cultured under normoxia or hypoxia, as evaluated by HIF1 alpha -responsive element luciferase reporter (HRE-luc) assay (n = 5). *P < 0.05 (one-way ANOVA followed by Dunnett’s post hoc test). Data are presented as mean ± SD. l mRNA level of hif1a in CON and Mapk7 CKO chondrocytes cultured under normoxia or hypoxia were measured by real-time quantitative PCR (n = 3). *P < 0.05 (one-way ANOVA followed by Dunnett’s post hoc test). Data are presented as mean ± SD. m Free ATP levels in CON and Mapk7 CKO chondrocytes cultured under normoxic or hypoxic conditions (n = 4). *P < 0.05 (one-way ANOVA followed by Dunnett’s post hoc test). Data are presented as mean ± SD. n Primary chondrocytes were transfected with WT-MAPK7 or DN-MAPK7 expression vector in the presence of CA-MEK5 expression vector. Cells were harvested 48 h after transfection and protein levels of HIF1 alpha , MAPK7 and p-MAPK7 were assayed by western blot Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32944217), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

TRF2 transcriptionally regulates p21 expression through promoter occupancy. (A) Quantitative ChIP using TRF2 antibody gives enriched binding of TRF2 on p21 promoter in HT1080 and MDA-MB-231 cells, IgG was used as isotypic antibody control; normalized with 10% input (data represented as mean ± SEM, for three replicates). (B,C) TRF2 represses p21 promoter activity. In luciferase assay, si-RNA-mediated silencing or TRF2 over expression resulted in increased (B, *p value < 0.05, Student’s t-test; data represented as mean ± SEM of three replicates) or reduction (C, *p value < 0.05, Student’s t-test; data represented as mean ± SEM of three replicates) in p21 promoter activity in HT1080 and MDA-MB-231 cells, respectively; over expression of TRF2 devoid of DNA binding [deletion of basic (delB), myb (delM) and both basic/myb (delB-delM) domains] resulted in partial or complete rescue of p21 promoter activity in HT1080 and MDA-MB-231 cells (data represented as mean ± SEM, for three replicates) (C). (D,E) HT1080 cells over expressing TRF2 show reduced p21 protein expression, bar graph shows the densitometry analysis of three different immunoblot replicates (*p value < 0.05, Student’s t-test; data represented as mean ± SEM of three replicates). (E) (Full images are shown in Supplementary Figure S5); TRF2 silencing results in increase in p21 protein, bar graph shows the densitometry analysis of three different immunoblots (*p value < 0.05, Student’s t-test; data represented as mean ± SEM of three replicates) (F); and mRNA expression in HT1080 cells (*p value < 0.05, Student’s t-test; data represented as mean ± SEM of three replicates, GAPDH used as internal control for real-time PCR). Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/s41598-017-11177-1), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: Goat Polyclonal AIF-1/Iba1 Antibody [NB100-1028] - AIF-1/Iba1 immunoreactivity in an FFPE section of mouse spleen. NB100-1028 was diluted to 1ug per mL and was left on tissue sections for 30m at room temperature. Secondary was horse anti goat HRP polymer. Image from a verified customer review.

Reactivity:

Hu, Mu, Rt, Ca, Eq, Fe, Po

Species Glossary

Reactivity: Hu, Mu, Rt, Ca, Eq, Fe, Po

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

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NB100-2323

IRE1 alpha [p Ser724] Antibody - BSA Free

(9 Reviews)

(318 Publications)

0.1 ml

NB100-2323

P-Ire1a activation correlates with WFS1 cell expression. Cryosections of retina from 12 month old Wfs1+/+ and Wfs1-/- mouse were immunostained with anti-P-Ire1a antibody (red). DAPI was used for staining of cell nuclei (blue). RPE, retinal pigment epithelium; ONL, outer nuclear layer; INL, inner nuclear layer, RGC, retinal ganglion cells. Scale bars=50 um. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0097222) licensed under a CC-BY license.

Hepatic ER stress markers with cachexia progression. ER stress markers Bip, IRE1a, ATF6p50 and CHOP were examined in the liver of non, pre and severely cachectic mice. (n = 6-8 per group, p < 0.05) Dotted line indicates levels of Non-cachectic mice. Non = Non-Cachectic Apc Min/+ Sev = severely cachectic Apc Min/+; * denotes significantly different from Non-cachectic Apc Min/+ Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0119888) licensed under a CC-BY license.

IRF-1 plays a central role in ER stress-mediated modulation of VCAM-1 expression by TGRL. HAEC were conditioned for 4 hr with TNFa (0.3 ng/ml) E: CHOP knockdown decreased TNFa-induced VCAM-1 expression. n=4. **P<0.01 vs. siCtrl+TNFa. Shown are representative blots from 3 independent experiments with similar results. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0078322) licensed under a CC-BY license.

Analysis using HRP conjugate of NB100-2323. Detection of phosphorylated IRE-1 alpha using NB100-2323. Lane 1: COS-7 untransfected Lane 2: COS-7 expressing wild-type IRE1 alpha Lane 3: COS-7 expressing kinase-dead IRE1 alpha. Theoretical molecular weight: 110 kDa.

<b>Biological Strategies Validation. </b>Analysis of anti-IRE1 alpha (pSer724) using Lot AN and AO of NB100-2323. HeLa cells were treated (+) or untreated (-) with 10 mM DTT for 60 min to activate the UPR. Total protein was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% BSA in TBST. The membrane was probed with 2.0 ug/ml antibody in 5% BSA, and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. Theoretical molecular weight: 110 kDa.

Regulation of the unfolded protein response by KLF15. Western analysis of UPR activity in WT versus KLF15-/- primary hepatocytes. Hepatocytes were isolated from standard chow-fed 4-month-old male WT and KLF15 -/- mice. Two individual experiments were performed in triplicate; each lane indicates a technical replicate. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0077851) licensed under a CC-BY license.

HeLa cell lysate, 20 ug. Antibody at 1:1000, 1% skim milk in TBST, overnight incubation at 4C. WB image submitted by a verified customer review.

<b>Biological Strategies Validation. </b>Detection in Min6 cells which were treated with different concentrations of glucose for 3 hours prior to lysates preparation.

IRE1 (pS724) was detected in immersion fixed paraffin-embedded sections of human spleen using Rabbit Anti-Human IRE1 (pS724) polyclonal Antibody (Catalog # NB100-2323) at 1:300 for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the perinuclear cytoplasm in splenocytes.

Hepatic ER Stress.Representative hepatic western blots of BiP, phosphorylated (Ser51), total EIF2 alpha and phosphorylated:total EIF2 alpha , phosphorylated (Ser724), total IRE1 alpha and phosphorylated:total IRE1 alpha , and CHOP (n = 9). Diets not sharing a common letter differ significantly from one another (P≤.05).

Morphine suppressed 6-OHDA-induced ER stress through activation of UPR. (A,B) Morphine induced UPR in SH-SY5Y cells. Protein levels of GRP78, p-IRE1 alpha , IRE1 alpha , p-PERK, PERK, ATF6, CHOP and Tubulin in SH-SY5Y cells were analyzed (A) and quantified (B) by western blot. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 vs. control.

Effect of cancer on ER stress markers.Bip1, IRE-1, ATF-6 p50 and CHOP expression in the liver of non—cachectic ApcMin/+ mice (N = 6 per group), compared to healthy C57BL/6 mice. Dotted line on the western blot indicates two different sections of the same gel. Values are expressed as Mean ± SE. * denotes significantly different from the healthy C57BL/6 mice as analyzed by a pre—planned t—test. p < 0.05.

T helper cells upregulate the IRE1a-XBP1 pathway during activation. c) Naive T helper cells and in vitro differentiated Th2 lymphocytes were analyzed for IRE1a mRNA expression by qRT-PCR (left panel), protein expression by Western blot (middle panel), and phosphorylated IRE1a (P-IRE1a) by Western blot (right panel). The density of Western blot bands from five independent experiments of IRE1a and three independent experiments of phospho-IRE1a were measured and displayed on top of each Western blot panel. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30355343), licensed under a CC-BY licence.

IER3IP1 suppression decreases the UPR activation. (B) Effect of IER3IP1 suppression on the IRE1alpha arm of the UPR. 4 days after IER3IP1 KD in MIN6 cells, phosphorylation of IRE1alpha, Bip, sXBP-1 and uXBP-were determined by Western blot. The loaded proteins in the two

CETP inhibition reduces endoplasmic reticulum stress. HAECs transfected with CETP or control scrambled siRNA. (B) Phosphorylated and total IRE (n = 3, ns). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33430172), licensed under a CC-BY licence.

Apoptosis mediated by Arl6ip5 interference is regulated by Chop. (a) Immunoblotting was used to detect the protein level for p-IRE1alpha, IRE1, Chop, Arl6ip5 and beta-actin in the UAMS-32 cell with NC-siRNA or Arl6ip5-siRNA treatment for 72 h. Representative results were shown from three independent experiments. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2014427), licensed under a CC-BY licence.

Activation of UPR signaling pathways after UIRI. e and f) The expression of p-IRE1 and t-IRE1 in mice kidneys were evaluated with western blotting and quantified. Data are expressed as means +/- SEM, n = 3 ~ 6 in each group. *P < 0.05, **P < 0.01, and ***P < 0.001, as compared with sham group Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35765067), licensed under a CC-BY licence.

Effect of Insig-1 overexpression on nSREBP-1c and proteins related to ER stress pathway in INS-1 cells. INS-1 and INS-1-Insig-1 cells were stimulated with 11.2 mM and 25.0 mM glucose for 0, 24 and 72 h. A) nSREBP-1c and p-IRE1alpha pathway of ER stress related proteins were detected by Western blot. GAPDH was used as control for protein loading. Equal amount of proteins (50 ug) were loaded. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/21843373), licensed under a CC-BY licence.

Protein expression of downstream markers of ER stress in islets from lean, obese and high-fat-diet-fed fZDF rats. Protein lysates from islets isolated from three age-matched fZDF rats (obese), HF-fZDF (HFD) rats and their heterozygous lean littermates (lean) were separated by SDS-PAGE and analysed by western blotting. As controls, lysates from the islets isolated from Wistar rats or MIN6 cells treated with 1 uM thapsigargin for 2 h or sorbitol for 1 h were run alongside. Proteins were detected using antisera against (a) Calreticulin, HYOU1, GRP78, p-IRE1alpha, p-eIF2alpha and CHOP and as protein loading control rpS6 and (b) p-JNK and total JNK1/2 as loading control. Western blots were quantified using Image-J software. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/nutd201335), licensed under a CC-BY licence.

ABT-263 induces ER stress and activation of the unfolded protein response. c) Kinetic analysis of ER stress proteins in Mel270 cells treated with ABT-263 5 uM in the absence or presence of qVD-OPh 20 uM. HSP90 is used as a loading control. Representative western blots are shown. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32337074), licensed under a CC-BY licence.

$Localization of the WT and mutant Wrn proteins in different subcellular fractions. (C) Protein levels of GRP78, phosphorylated IREalpha, and beta-actin (top panels) and of total IREalpha, and beta-actin (bottom panels) in the spleen of three animals of each genotype. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26447695), licensed under a CC-BY licence.$

Glucocorticoids suppress NIX expression independent from PINK1-parkin pathway.a SH-SY5Y cells were incubated with cortisol for 12 h and mRNA expressions were analyzed by real-time PCR. n = 5. b, c Hippocampal neurons and SH-SY5Y cells were treated with corticosterone for 24 h and with cortisol for various time, respectively. Expression of NIX, PTEN-induced kinase 1 (PINK1), and BCL2 interacting protein 3 (BNIP3) were detected via western blot. n = 5. d, e Hippocampal neurons and SH-SY5Y cells were treated with corticosterone and cortisol, respectively, for 24 h. NIX (green), TOMM20 (red), and DAPI (nuclear counterstaining, blue) were visualized. Scale bars, 20 μm (magnification, ×1000). n = 5. f Nontargeting (NT) or PARK2 siRNA was transfected to SH-SY5Y cells for 24 prior to cortisol for 24 h. NIX was co-immunoprecipitated with parkin. Parkin levels in immunoprecipitated samples were quantified. n = 5. g, h Both hippocampal neurons and SH-SY5Y cells were transfected with mitochondria-targeted Keima (mt-Keima)-Red-Parkin for 24 h prior to corticosterone and cortisol for 24 h, respectively. Western blot was performed. n = 5. i–l Both hippocampal neurons and SH-SY5Y cells were pretreated with phorbol 12-myristate 13-acetate (PMA, 10 nM) for 30 min before corticosterone and cortisol for 24 h, respectively. i, j Hippocampal neurons and SH-SY5Y cells were transfected with mt-Keima at DIV 5 and 48 h prior to treatment, respectively. DAPI was used for nuclear counterstaining (blue). Ratio of red to sum of red and green was quantified. Scale bars, 20 μm (magnification, ×1000). n = 5. k, l TOMM20 levels were detected by western blot. n = 5. m SH-SY5Y cells were transfected with pcDNA3.1/c-eGFP or pcDNA3.1/NIX-c-eGFP vector for 24 h prior to cortisol for 24 h. TOMM20 levels were detected by western blot. n = 5. All blots and immunofluorescence images are representative. n = 5 from independent experiments with two technical replicates each. Quantitative data are presented as a mean ± S.E.M. The representative images were acquired by SRRF imaging. Two-sided unpaired student’s t test: Figs. 3a, b, d, e. Two-sided one-way ANOVA: Fig. 3c. Two-sided two-way ANOVA: Figs. 3f–m. ** indicates p < 0.01 versus control. ## indicates p < 0.01 versus corticosterone in hippocampal neurons and cortisol in SH-SY5Y. Data are provided as a Source data file. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/33473105), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Expression analysis of mouse Fchsd1 and Fchsd2 in different tissues.(A) Total RNA from postnatal day 5 mouse tissues was extracted and used as template for reverse transcription. PCR was performed using this cDNA as template. Upper panel, Fchsd1 mRNA is expressed more abundantly in nervous tissues. Middle panel, Fchsd2 mRNA is expressed ubiquitously in all tissues examined. Lower panel, beta -actin specific primers were used as the RT-PCR template control. (B) Total proteins of postnatal day 5 mouse cochlea and vestibula were extracted and separated by PAGE and detected with antibodies against FCHSD1(Novus), FCHSD2, or SNX9. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/23437151), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

LAIR-1 inhibits Glut1-related glucose uptake in OS cells. a Heatmap showing the levels of differentially expressed mRNAs. b Top 20 KEGG pathway annotation categories for target gene functions of predicted mRNAs. c Selected significantly differentially expressed mRNA-related to EMT in RNA-seq data between two groups, ***P < 0.001. d qPCR validation of differentially expressed EMT-related genes in LV-NC and LV-LAIR-1-overexpressing OS cells, **P < 0.01. e Glut1 expression analyzed by western blotting. f Immunofluorescence staining of Glut1 in the LV-LAIR-1-overexpressing OS cells. Scale bar = 50 μm. Data were obtained from at least two independent experiments. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32563267), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Phase contrast microscopy images of HFA and brain tumor cells and immunofluorescence images of cultured primary brain tumor cells, AA and HFA.HFA were derived from three different foetal brains. (A) 18-week foetus (10× magnification); (B) 19-week foetus, first week in culture (10× magnification); (C) 19-week foetus, day 5 in culture (20× magnification); (D) Primary GBM after 2 trypsinisations (20x); (E) Recurrent GBM (20x); (F) Secondary GBM after 3 trypsinisations (10x); (G) OII 14 days in culture (10x); (H) AII 13 days in culture (10x); (I) Negative control for NGS merged with DAPI; (J) Negative control for secondary antibodies merged with DAPI; (K) GBM cells derived from one patient – GFAP (DAKO) merged with DAPI and CD68 (zoomed: 2.4× magnification); (L) primary brain tumor cells from one patient (male, 40 years old) (passaged three times) - GFAP (Novocastra) merged with DAPI; (K and L) GFAP (green) was used as a GBM marker and CD68 (red) was used as a marker for microglia. (M) AA from one Female, 61 years old; (passaged twice) - GFAP (Novocastra) merged with DAPI; (N) AA from one male 40 years old; (passaged once) - GFAP (Sigma) merged with DAPI and CD68 (Abcam); (M and N) GFAP (green) was used as an astrocyte marker and CD68 (red) and CD11b (green) were used as markers for microglia; (O) HFA from one 17-week- old foetus (passaged once) - GFAP (Novocastra) merged with DAPI and CD11b (Novus); (P) HFA from an 18-week- old foetus (passaged once) - GFAP (DAKO) merged with DAPI and CD68 (zoomed: 2.1× magnification); (O and P) GFAP (green and red) was used as an astrocyte marker and CD68 (red) and CD11b (green) were used as markers for microglia. Nuclei indicated by DAPI (blue) in all images. Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.pone.0112945), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Regulation of FOXC1 on TLR3/4 expression in mice subjected to myocardial ischaemia (MI). The siRNA against FOXC1 (si‐FOXC1) or FOXC1 adenovirus (Ad‐FOXC1) was injected into the left ventricle just after LAD ligation to generate FOXC1 knockdown (A) or FOXC1 overexpression (B), and the negative control (NC) siRNA and Ad‐GFP served as control, respectively. The mRNA (upper panel) and protein (middle and lower panel) levels of TLR3/4 were determined after 2 wk. Data are means ± SEM aP < .05, AP < .01 vs respective MI Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31517441), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Analysis of protein levels of genes with increased transcription in hypoxia, IOX2 and VH032.HIF targets were increased in hypoxia, VH298 and FG-4592. 0.05% DMSO (vehicle control), 1% O2 (hypoxia), 100 µM VH298 and 50 µM FG-4592 were introduced to (A) HeLa or HFF for 24 hours and (B) HeLa for indicated time. Protein levels were analysed by immunoblotting using antibodies against indicated proteins, with beta -Actin as loading control. The blots shown are representative of three independent experiments. * indicates longer exposure. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30801039), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Dr, Gt, Ma, Pm, Pm, Rb, SyHa

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Dr, Gt, Ma, Pm, Pm, Rb, SyHa

Applications:

, , , , , , , ,

Applications: , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-2323AF350
Alexa Fluor 405	NB100-2323AF405
Alexa Fluor 488	NB100-2323AF488
Alexa Fluor 532	NB100-2323AF532
Alexa Fluor 594	NB100-2323AF594
Alexa Fluor 647	NB100-2323AF647
Alexa Fluor 700	NB100-2323AF700
Alexa Fluor 750	NB100-2323AF750
Allophycocyanin	NB100-2323APC
Biotin	NB100-2323B
DyLight 350	NB100-2323UV
DyLight 405	NB100-2323V
DyLight 488	NB100-2323G
DyLight 550	NB100-2323R
DyLight 594	NB100-2323DL594
DyLight 650	NB100-2323C
DyLight 680	NB100-2323FR
DyLight 755	NB100-2323IR
FITC	NB100-2323F
HRP	NB100-2323H
Janelia Fluor 549	NB100-2323JF549
Janelia Fluor 646	NB100-2323JF646
PE	NB100-2323PE
PerCP	NB100-2323PCP
mFluor Violet 450 SE	NB100-2323MFV450
mFluor Violet 500 SE	NB100-2323MFV500
mFluor Violet 610 SE	NB100-2323MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-2323SS

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NB100-449

HIF-1 alpha Antibody

(28 Reviews)

(286 Publications)

0.1 ml

NB100-449

Simple Western: HIF-1 alpha Antibody [NB100-449] - Simple Western lane view shows a specific band for HIF-1 alpha in 0.5 mg/ml of hypoxic HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Immunohistochemistry: HIF-1 alpha Antibody [NB100-449] - Mouse Brain, Neurons 40X

Immunohistochemistry: HIF-1 alpha Antibody [NB100-449] - Detection of human HIF1-alpha by immunohistochemistry. Sample: FFPE section of renal cell carcinoma. Antibody: Affinity purified rabbit anti-HIF1-alpha antibody (NB110-58773). Detection: DAB

<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody [NB100-449] - Detection of Human HIF-1 alpha by Western Blot and Immunoprecipitation. Samples: Whole cell lysate (5, 15 and 50 ug for WB; 1 mg for IP, 20% of IP loaded) from HeLa cells that were either treated with cobalt chloride (+; 200 uM) or mock treated (-). Antibodies: Affinity purified rabbit anti-HIF1 alpha antibody used for WB at 0.1 ug/mL(A) and 1 ug/mL (B) and used for IP at 3 ug/mg lysate. HIF-1 alpha was also immunoprecipitated by a previous lot of this antibody. Detection: Chemiluminescence with exposure times of 30 seconds (A) and 10 seconds (B).

Western Blot: HIF-1 alpha Antibody [NB100-449] - Blot showing the effect of hypoxia on the protein expression levels of the purine biosynthetic enzymes. HIF-1 alpha is stabilized in hypoxia as expected, and no significant increase in the purine enzymes was detected between normoxic (21% oxygen) and hypoxic (1% oxygen) growth conditions. The positions of molecular markers surrounding each band of interest are shown for each blot. ADSL (NBP2-03107), ATIC (NBP2-01941), FGAMS (NBP1-84691), GART (H00002618-M01), HIF-1a (NB100-449), PAICS (NBP2-02817), PPAT (NBP2-02056). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32439803/) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody [NB100-449] - Murine primary bone marrow derived macrophages stained with HIF1-alpha antibody (red). Nuclei were counterstained with DAPI (blue). Image from verified customer review.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody [NB100-449] - Analysis of microvascular density and HIF-1alpha activity. Low, homogenous expression of HIF-1alpha was detected in the tumors of NNK treated mice (40X, scale bar = 25 um). Image collected and cropped by CiteAb from the following publication (//molecular-cancer.biomedcentral.com/articles/10.1186/1476-4598-11-4), licensed under a CC-BY license.

Flow Cytometry: HIF-1 alpha Antibody [NB100-449] - HeLa cells were treated for 15 hrs with 200 uM CoCl2, fixed in PFA, and permeabilized in 90% MeOH. 1 X 10^6 cells were stained with 0.125 ug anti-HIF-1 alpha and secondary FITC-conjugated goat anti-rabbit (in a 150 uL reaction). Black: treated, anti-KLH control IgG; Red: untreated, anti-HIF-1 alpha; Blue: treated, anti-HIF-1 alpha.

<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody [NB100-449] - Analysis of HIF-1 alpha in human myeloma cell lysate using anti-HIF-1 alpha. Cells were untreated or treated with IGF-1, IL-6 or CoCl2. Image from verified customer review.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Detection of HIF-1 alpha in a hypoxic sample. Lane 1: CoCl2 treated Cos-7 nuclear extract (hypoxic). Lane 2: Untreated Cos-7 nuclear extract (normoxic).

Western Blot: HIF-1 alpha Antibody [NB100-449] - Detection of mouse HIF-1 alpha on hypoxia treated MEFs

<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody [NB100-449] - Samples: Whole cell lysate (5, 15 and 50 ug) from HeLa cells that were treated with cobalt chloride (+; 200 uM) or mock treated (-). Antibodies: Affinity purified rabbit anti-HIF1 alpha antibody NB100-449 used for WB at 0.1 ug/ml. Detection: Chemiluminescence with exposure times of 30 seconds.

Western Blot: HIF-1 alpha Antibody [NB100-449] - BMDM were seeded at 0.5x10^6 overnight. Cells were treated with 10 ng/ml LPS for 24 hrs. Image from verified customer review.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Analysis of total protein using anti-HIF-1 alpha, -CAIX, -ISCU antibodies and VDACs antibody (PMID: 29596470).

Western Blot: HIF-1 alpha Antibody [NB100-449] - Analysis of total protein using anti-HIF-1 alpha, -ISCU, -FXN antibodies. CoCl2 was used to treat or not HeLa cells for 2 days (PMID: 29596470).

Western Blot: HIF-1 alpha Antibody [NB100-449] - Detection of human HIF1 alpha by western blot. Samples: Whole cell lysate (15 and 50 ug) from HEK293T cells that were either treated with 200 uM cobalt chloride (+) or mock treated (-). Antibodies: Affinity purified rabbit anti-HIF1 alpha antibody NB100-449 used for WB at 0.1 ug/ml. Detection: Chemiluminescence with exposure times of 3 minutes.

Western Blot: HIF-1 alpha Antibody [NB100-449] - 3T3-L1 mouse embryonic fibroblast adipose-like cell line. Antibody at 1:2000. WB image submitted by a verified customer review.

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody [NB100-449] - Formaldehyde-fixed asynchronous HeLa cells.

Immunoprecipitation: HIF-1 alpha Antibody [NB100-449] - HIF-1 alpha analysis in HEK293 cells. Image from verified customer review.

Immunoprecipitation: HIF-1 alpha Antibody [NB100-449] - Detection of human HIF1 alpha by western blot of immunoprecipitates. Samples: Whole cell lysate (1 mg for IP, 20% of IP loaded) from HEK293T cells that were treated with 200 uM cobalt chloride. Antibodies: Affinity purified rabbit anti-HIF1 alpha antibody NB100-449 used for WB at 1 ug/ml and for IP at 6 ug/mg lysate. Detection: Chemiluminescence with exposure times of 3 minutes.

beta -Glucan size affects the requirement for reactive oxygen species (ROS) in IL-1 beta induction.(B) Human mDCs from healthy donors or chronic granulomatous disease (CGD) patients were stimulated with curdlan or glucan-mp for 8 h. HIF-1 alpha and pro-IL-1 beta protein expression measured by immunoblot (n = 2 donors).

Hypoxia downregulates CFTR expression. (e) Western blot showing the expression changes of HIF-1alpha and EMT markers induced by hypoxia in MDCK cells, quantification analysis is shown in the lower panel, *p < 0.05; (Full-length blot is shown in Supplementary Figure S7d.). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28701694), licensed under a CC-BY licence.

BCAT1 expression and inhibition in human macrophages. (e) Western blot analysis of IRG1, IL-1beta, HIF-1alpha and beta-actin (ACTB) in control (untreated), ERG240-treated (20 mM, 3 h), LPS treated (100 ng ml-1, 3 h) and LPS+ERG240 treated (LPS, 100 ng ml-1; ERG240, 20 mM for 3 h) hMDMs. The experiment is representative of three independent experiments using n=3 healthy donor hMDMs each. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms16040), licensed under a CC-BY licence.

Elevated PIK3/AKT/HIF-1alpha pathways in KL SqCC tumours. (c) IHC analysis (top) and quantification (bottom) of p63, p-AKT, p-S6, p-4EBP1, HIF-1alpha and GLUT1 in KL tumours (n=6 each group). Two-tailed t-test, ****P<0.0001, ***P<0.001, **P<0.01. Scale bar, 50 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28548087), licensed under a CC-BY licence.

Adenosine mediates increase in pro-IL-1beta via a HIF-1alpha-dependent pathway. (h) LPS primed PECs obtained from HIF-1alpha-cKO and control mice were treated with or without NECA at different time points as indicated followed by pulsing with ATP. Cell lysates were collected after ATP pulsing. HIF-1alpha, pro-caspase-1, Caspaspe-1 and pro-IL-1beta was measured in cell supernatant by western blot. Immunoblots shown are representative results from at least three independent experiments. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24352507), licensed under a CC-BY licence.

Morphological alterations induced by DMOG are HIF-1alpha dependent. (A) glEND.2 cells were stably transfected with shGFP, shHIF-1alpha or shHIF-2alpha. The clones were cultured under normoxic (N) or hypoxic (H, 1% O2) conditions, or treated with DMOG (D, 1 mM) for 6 h. 30 ug protein of cellular protein were loaded for the detection of HIF-1alpha, and nuclear extracts (20 ug protein) were used for the detection of HIF-2alpha. Image collected and cropped by CiteAb from the following publication (//biosignaling.biomedcentral.com/articles/10.1186/1478-811X-11-80), licensed under a CC-BY licence.

Pyruvate kinase isoform M2 (PKM2) and hypoxia-inducible factor 1alpha (Hif-1alpha) bind directly to the promoter region of PDL1 in primary murine BMDM cells. BMDM cells were treated with TEPP-46 at 50 uM for 1 h prior to stimulation with LPS (100 ng/ml, 24 h) (C) Sequential ChIP assays measuring simultaneous endogenous binding of PKM2 and Hif-1alpha to chromatin in response to LPS (100 ng/ml, 24 h) +/-TEPP-46 (pretreatment using 50 uM, 60 min). ChIP data are calculated as percent of input, error bar represents mean +/- SEM, and statistics are performed as two-tailed unpaired t-test *P < 0.05, **P < 0.01, and ***P < 0.001. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29081778), licensed under a CC-BY licence.

Changes in Hepatic Oxygen Saturation and Hypoxia-related Protein Expression. (C) In the Western blot analysis, the experimental group was relatively higher expression of hypoxia-inducible factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) protein than the control group at eight weeks. The full-length blots with these antibodies were presented in supplementary Figure S1. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29079853), licensed under a CC-BY licence.

Effects of cell stress on VEGF-A and HIF1alpha expression in Muller cells and Y79 photoreceptors. (B) Western blots for HIF1alpha using cellular proteins, n = 4/group, ** p < 0.01. For MIO-M1 Muller cells: Grey bars: 5 mM glucose. Black bars: 25 mM glucose. For Y79 photoreceptors: Grey bars: 11 mM glucose. Black bars: 25 mM glucose. NS, not significant; LG, low glucose; HG, high glucose. Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/1422-0067/18/3/533), licensed under a CC-BY licence.

Hypoxic conditions upregulate CNN3 in BeWo cells.BeWo cells were cultured either under normoxic or under hypoxic conditions for 24hr. (A) Total protein lysates were examined with the Western Blot technique to detect CNN3 protein levels. For normalization, HPRT was stained on the same membrane. As hypoxia marker, HIF-1 alpha was detected as well. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25050546), licensed under a CC-BY licence.

Phenylhydrazine administration results in hypoxia in retinal vessels and in HIF-1alpha and HIF-2alpha stabilization in peripheral blood leukocytes. (B) Representative analysis of HIFs by western blot: 50ug of protein from total cell lysate of blood leukocytes isolated from mice treated with PHZ compared to untreated. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28112274), licensed under a CC-BY licence.

Western Blot: Rabbit Polyclonal HIF-1 alpha Antibody - Analysis of HIF-1 alpha antibody on mouse liver nuclear extracts. Image from a verified customer review.

Improved survival and blood–brain barrier function post pneumococcal infection in mice by anti-HIF-1 alpha treatment using echinomycin. a Schematic depicting treatment protocol of mice infected with S. pneumoniae (D39). Mice were infected i.p., followed by anti-HIF-1 alpha treatment with echinomycin or just vehicle every 12 h starting 4 h post-infection (blue tick marks). Clinical scores were taken every 4 h starting 16 h post-infection (red line with tick marks) and study terminated at 48 h post-infection and brain tissues collected upon animal death or at the end point when the animals were sacrificed. b Clinical scores indicate improved symptoms in echinomycin-treated mice compared to vehicle-treated animals starting 16 h post-infection that were progression free (below score 1) up to 24 h (mean ± SEM at each time, N = 10/group, ****p < 0.0001, **p < 0.01, *p < 0.05 by 2-tailed unpaired non-parametric Mann–Whitney t test at each time point. c Echinomycin-treated mice showed significantly improved overall survival percentage by Kaplan–Meier analysis with the median survival at 32 h compared to 25 h in vehicle-treated mice (**p < 0.01, log-rank test, N = 10/group). d Kaplan–Meier analysis also indicated a significant improvement in progression-free survival in echinomycin-treated animals (median survival 25 h) with no clinical symptoms up to 24 h (b) compared to the vehicle group (median survival 17 h; ***p < 0.001, log-rank test, N = 10/group). e Immunofluorescence staining for HIF-1 alpha , BBB permeability and junctional markers in the echinomycin and vehicle groups at the survival end point. Left panel shows that echinomycin treatment leads to a reduction in HIF-1 alpha -positive nuclei including in ECs co-stained for podocalyxin, a vascular marker which was unchanged by the treatment. Middle panel displays reduced vascular permeability to fibrinogen in the echinomycin group as indicated by stronger intravascular signal compared to the vehicle-treated mice. Increased expression of tight junction proteins—occludin (middle panel) and claudin-5 (right top panel)—in echinomycin-treated mice indicate improved BBB function. Tight junction-associated ZO-1, adherens junction marker VE-Cadherin, and endothelial cell adhesion molecule CD31 were unchanged (middle, bottom right panel). There was also no difference in S. pneumoniae (Spn) staining (right top panel) between the two groups. Scale bar 10 μm. f Quantification of the staining from e utilizing four images per animal in the cortex region shows a significant reduction in HIF-1 alpha -positive cell number, whereas the total cell number or EC cell number was unchanged (left panel). As observed in e, the middle panel for quantification (arbitrary units—a.u.) of fibrinogen leakage shows significantly increased vascular staining for fibrinogen, supported by significantly increased expression of occludin, claudin-5, whereas other EC junction markers were unchanged. S. pneumoniae numbers in the vessels (v-Spn) or those transmigrated into brain parenchyma (b-Spn) were also unchanged. (mean ± SEM, N = 7–10/group indicated by the corresponding number of dots, ****p < 0.0001, **p < 0.01, *p < 0.05 by 2-tailed unpaired Student’s t test) Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32529267), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Compound MI-743 causes intracellular 8-oxo-dG accumulation and DNA damage.a MGC-803, HGC-27 and GES-1 cells were treated with DMSO or 5 µM MI-743 for 48 h. Intracellular 8-oxo-dG was stained with Cy3-conjugated avidin. DNA was counterstained with 4,6-diamidino-2-phenylindole (DAPI). Images were acquired at ×100 magnification by a Nikon Eclipse TE 2000-S fluorescence microscope. At least three independent experiments were performed for each group. b MGC-803, HGC-27 and GES-1 cells were treated with DMSO or 10 µM MI-743 for 48 h and run in alkaline comet assay. Pictures were originally captured at ×40 magnification. H2O2 was used as positive control. c Tail moment was calculated by CometScore software. Three individual experiments were performed for each group. d–f Western blot analysis of the protein levels of MTH1, MUTYH, OGG1, p21, ATMpS1981 and p53pS15 in MGC-803, HGC-27 and GES-1 cells, treated with increasing concentrations of MI-743 (0, 1, 2, 4, 8 and 12 µM). g, h Densitometry shows relative protein expression normalized for GAPDH. Data are representative of three independent experiments. i–k Western Blot analysis of the protein levels of MTH1, MUTYH, OGG1, p21, ATMpS1981 and p53pS15 of protein lysates, isolated from MGC-803, HGC-27 and GES-1 cells, which were treated with 12 µM MI-743 for 0, 12, 24, 36 and 48 h. l, m Densitometry shows relative protein expression normalized for GAPDH. Data are presented as means ± SD. Three individual experiments were performed for each group. *P < 0.05, **P < 0.01, ***P < 0.001 as compared with the controls Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31164636), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Successful knockdown of HIF-2 alpha in both 786-O and A498 renal cell lines. (A) Lysates from 786-O cells stably transfected with VHL (VHL +), parental 786-O cells (786-O), and 786-O cells infected with an empty retrovirus (pSuperRetro) or with retroviruses encoding single HIF-2 alpha shRNAs (HIF2 alpha shRNA 1; HIF2 alpha shRNA 2) or doubly infected with both shRNAs (HIF2 alpha shRNA 1+2) were immunoblotted for HIF-2 alpha (top panel), GLUT1 (upper middle panel) and VHL (lower middle panel). Bottom panel is an alpha tubulin immunoblot to demonstrate equal protein loading (25 μg) in each lane. (B) Lysates from A498 cells stably transfected with VHL (VHL +), parental A498 cells (A498), and A498 cells infected with an empty retrovirus (pSuperRetro) or doubly infected with both HIF-2 alpha shRNAs (HIF2 alpha shRNA 1+2) were immunoblotted as in (A). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/17598890), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

IQGAP1 induces the epithelial-to-mesenchymal transition, invasiveness, and proliferation of endometrial cancer (EC) cellsA. Reverse transcription quantitative PCR (qPCR) analysis of IQGAP1 mRNA in the immortalized human endometrial cell line EM and the EC cells HEC-1, HEC-50, and HEC-50-HI (HI). The results are presented as the fold-change in expression compared to the EM cells. B. Western blot analysis of the IQGAP1 protein in EC cells. C, E. The expression of IQGAP1, E-cadherin, and N-cadherin proteins in HI cells transfected with control (Ctr) or IQGAP1 siRNA (C) and in HEC-1 cells expressing either the control or IQGAP1 vector (E). D, F. Phase-contrast microscopy shows the morphology of HI cells transfected with control or IQGAP1 siRNA (D) and HEC-1 cells transfected with the control or IQGAP1 vector (F). G-I. Detection of migration (G), invasion (H), and proliferation (I) in HI and HEC-1 cells after the indicated transfection. J, K. qPCR analysis of ZO-1, CK-18, and Vimentin expression in HI (J) and HEC-1 (K) cells, transfected as indicated. L. Representative images from the invasion assays. Image collected and cropped by CiteAb from the following open publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.7754), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

FDFT1 inhibits glycolysis through suppressing the AKT-mTOR-HIF1 alpha pathway in CRC.aFDFT1 overexpression reduced glucose uptake in CT26 cells. bFDFT1 overexpression decreased lactate production via glycolysis in CT26 cells. c ECAR was reduced when FDFT1 was overexpressed in CT26 cells. d OCR was increased when FDFT1 was overexpressed in CT26 cells. eFDFT1 knockdown increased glucose uptake in CT26 cells. fFDFT1 knockdown increased lactate production via glycolysis in CT26 cells. g ECAR was increased when FDFT1 was knocked down in CT26 cells. h OCR was decreased when FDFT1 was knocked down in CT26 cells. i, mFDFT1 overexpression inhibited the protein and mRNA expression of mTOR-targeted glycolytic enzymes, including GLUT1, HK2, PGK1, GPI, and LDHA, in CT26 cells. j, mFDFT1 knockdown increased the protein and mRNA expression of mTOR-targeted glycolytic enzymes, including GLUT1, HK2, PGK1, GPI, and LDHA, in CT26 cells. k, nFDFT1 overexpression decreased the protein and mRNA expression of AKT, mTOR, and HIF1 alpha . l, nFDFT1 knockdown increased the protein and mRNA expression of AKT, mTOR, and HIF1 alpha . o Photograph of dissected tumors (the first line: normal diet, the second line: FMD + glucose, the third line: FMD, n = 5). p The tumor volumes were measured every 2 days after the 13th day. The FMD + glucose group can reverse the tumor growth inhibition induced by the FMD (n = 5; ns: P = 0.1838; P = 0.0001). q The protein level of FDFT1 and mTOR in dissected tumor samples from normal diet group, FMD group and FMD + glucose group was measured by western blotting. r The glucose level in these three groups. Error bars, mean ± SD, the data are from three independent experiments. Two-sided t tests. *P < 0.05, **P < 0.01, ***P < 0.001, compared with the control group (or normal diet group). #P < 0.05, ##P < 0.01. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32313017), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Effect of MLL-HCF1 interaction on histone methylation and Nrf2 expression in SNUC5/5-FUR cellsA. HCF1 expression was determined by Western blotting. *Significantly different from SNUC5 cells (p<0.05). B. Interaction between MLL and HCF1was examined by immunoprecipitation analysis using anti-MLL and anti-HCF1 antibodies followed by Western blotting with anti-HCF1 and anti-MLL antibodies. *Significantly different from SNUC5 cells (p<0.05). C. Interaction between MLL and HCF1was assessed by PLA. Each green spot represents for a single interaction (MLL and HCF1) and DNA was stained with DAPI. *Significantly different from SNUC5 cells (p<0.05). The cells were transfected with non-targeting siRNA (siControl) or siHCF1 RNA for 24 h. D. Expression pattern of MLL mRNA in SNUC5/5-FUR cells was determined by RT-PCR analysis. *Significantly different from siControl-transfected cells (p<0.05). E. Expression pattern of HCF1, MLL, and Nrf2 in SNUC5/5-FUR cells was determined by Western blot analysis. *Significantly different from siControl-transfected cells (p<0.05). Image collected and cropped by CiteAb from the following open publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.9745), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Induction of ubiquitin/proteasome-dependent DSBs in CPT-treated quiescent WI38 hTERT cells. (A–F) Serum-starved cells were treated with DMSO (1 h) or with MG132 (50 μM, 1 h), lactacystin (10 μM, 1 h), bortezomib (1 μM, 4 h), G5 (1.5 μM, 0.5 h) or Pyr-41 (9 μM, 0.5 h) before the addition of DMSO (untreated) or 25 μM CPT for 1 h and then co-stained for gamma H2AX (green) and 53BP1 (red) or analyzed by Western blot. ‘-’ in panels C and F means cells treated with DMSO. (A and D) Representative pictures. Nuclear contours, identified by DAPI staining (not shown), are indicated by dashed lines. Bars: 10 μm. (B and E) Number of gamma H2AX foci per nucleus from two independent experiments (147–153 nuclei were analyzed for each treatment). ****P < 0.0001. (C and F) Western blot of gamma H2AX. alpha Tubulin: loading control. Dashed lines indicate that intervening wells have been spliced out. The top panels show quantification of gamma H2AX normalized to alpha Tubulin (means ± SEM, n = 4 in panel C, n = 3 in panel F). ***P < 0.001; **P < 0.01. (G and H) Detection of DSBs by neutral Comet assays in serum-starved cells treated with DMSO or MG132 (25 μM) for 1 h before the addition of DMSO (untreated) or CPT for 1 h (7.5 μM for experiments (Exp) I and II; 5 and 7.5 μM for Exp III). (G) Representative pictures of nuclei from Exp I. (H) Quantification of neutral Comet tail moments for three independent experiments (95–133 nuclei were analyzed for each treatment in each experiment). ***P < 0.001; ****P < 0.0001. The untreated and CPT data from Exp I are from the same experiment as that of Supplementary Figure S3D. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/26578593), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$S100A7 is secreted and acts as a chemotactic factor of cell migration and invasionA. Cells were cultured in serum-free medium for 2 days, then the medium was collected, fractioned, followed by Western Blot analysis. B&C. Transwell migration and Matrigel invasion assays were performed in C33A and SiHa cells. Cell suspension was placed into the upper chamber in 0.1 ml of DMEM serum-free medium, and conditioned medium from S100A7-overexpressed C33A and SiHa cells and their corresponding control cells was placed in the lower chamber as a chemoattractant. The stained cells were manually counted from 4 randomly selected fields and normalized with cell proliferation [2-sided t test; *P < 0.05; ***P < 0.001]. Representative image and quantitative results of cell migration and invasion were shown (B. C33A cells; C. SiHa cells; ×10, bars:100μm). Image collected and cropped by CiteAb from the following open publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.15329), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Increased expression of HIF alpha and/or Twist in A549 and H441 cells induced by the inhibition of PIMT and Thapsigargin(A) Immunoblotting of Slug, ZEB1, Snail1, Twist, and HIF1 alpha in A549 sh-PIMT and sh-control cells. (B, C) Immunoblotting and relative intensity of HIF1 alpha in A549 cells treated with Tg. (D) Immunoblotting of Slug, ZEB1, Snail1, Twist, and HIF1 alpha in si-control cells and si-PIMT H441 cells. (E, F) Immunoblotting and relative intensity of HIF1 alpha in H441 cells treated with Tg. #1 and #2 indicates si-RNA of J-010000-05-0002 and J-010000-07-0002, respectively. Image collected and cropped by CiteAb from the following open publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.24324), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Characterization of CIP2A expression in a panel of normal human melanocytes (NHM) and melanoma cell lines. (A) Immunoblot analysis and (B) quantification of protein expression levels of CIP2A (C) CIP2A mRNA expression measured by qRT-PCR. NHM1 line was used as calibrator. (D) Cytoplasmic and nuclear expression of CIP2A in normal melanocytes, primary and metastatic melanoma cell lines assessed by immunoblot analysis and (E) by immunohistochemical staining. CIP2A, cancerous inhibitor of protein phosphatase 2A. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/25663244), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Role of inflammasome in glucose fluctuation-induced myocardial fibrosis. (A) Immunofluorescence staining of intracellular NLRP3–ASC interaction examined by confocal microscopy. The NRCFs were labeled with anti-NLRP3 (red), anti-ASC (green), and DAPI (blue) (n = 3 per group). (B–G) After siRNA targeting NLRP3 transfection, the protein expressions of NLRP3, cleaved caspase-1 and collagen I were measured by western blot (n = 3 per group). (H) Representative images of immunofluorescence staining of intracellular alpha -SMA in the NG, HG, GF, and GF + siRNA groups (n = 3 per group). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35592403), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Ca, Ch, Gt, Pm, Pm, Bv

Species Glossary

Reactivity: Hu, Mu, Rt, Ca, Ch, Gt, Pm, Pm, Bv

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

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NB100-417

Carbonic Anhydrase IX/CA9 Antibody - BSA Free

(7 Reviews)

(271 Publications)

0.1 ml

NB100-417

CA9 expressed in hypoxic regions as assessed by pimonidazole staining (A), but also observed in non-pimonidazole areas (B). Most of CA9 expressed in pimonidazole positive regions (C). Proliferation (BrdUrd) & apoptosis (caspase-3) in relation to CA9 expression shown in figure D & E. Red, CA9; Green, pimonidazole (A-B), BrdUrd (D) or caspase-3 (E); Yellow, overlap of CA9 (red) & pimonidazole (green); Light blue, vessels. Magnification 100x. Scale bars represent 100 um. Closed circles represent CA9 expression in pimonidazole positive regions; open circles represent CA9 expression in pimonidazole -ve regions. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0108068), licensed under a CC-BY license.

<b>Orthogonal Strategies Validation. </b>Formalin-fixed paraffin-embedded tissue sections of human stomach were probed for Carbonic Anhydrase IX/CA9 mRNA (ACD RNAScope Probe, catalog # 559348; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using Rabbit Polyclonal (Novus catalog # NB100-417) at 1:1000 dilution with overnight incubation at 4 degrees Celsius followed by incubation with anti-rabbit IgG VisUCyte HRP Polymer Antibody (Catalog # VC003) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to glandular cells.

Analysis on various human cell lysates. Specific bands were detected for Carbonic Anhydrase IX/CA9 in HEK-293 and A498 cell lines at a molecular weight of approximately 50 kDa. WB image submitted by a verified customer review.

Immunohistochemical representative microphotographs representing the HIF-1alpha, GLUT-1, and CAIX expression in endometrial cancer according to FIGO classification (IA, IB, II, IIIA, IIIC, and IV). Primary objective magnification 20x. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/bmri/2014/616850/) licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>Hypoxias impact on proteins of the mitochondrial ISC assembly machinery. (A) Immunoblotting analyzed the total protein extracts from HeLa cells grown in normoxia (Nx, 21% O2) or hypoxia (Hx, 1% O2) conditions using VDACs poly antibody and anti-CAIX, -ISCU, -FXN, -NFS1, -HSC20 antibodies. Beta-Actin was used as loading control. Citation: Ferecatu I, Canal F, Fabbri L, Mazure NM, Bouton C, Golinelli-Cohen M-P (2018) Dysfunction in the mitochondrial Fe-S assembly machinery leads to formation of the chemoresistant truncated VDAC1 isoform without HIF-1 alpha activation. PLoS ONE 13(3): e0194782.

<b>Biological Strategies Validation. </b>Analysis using the DyLight 488 conjugate of NB100-417. Staining of Carbonic Anhydrase IX (red) in human glioma U87 cells. DAPI counterstains nuclei (blue). ICC/IF image submitted by a verified customer review.

Simple Western lane view shows a specific band for CAIX in 0.1 mg/mL of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

An intracellular stain was performed on U-87 MG Cells with NB100-417 and a matched isotype control. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG APC-conjugated Secondary Antibody, (R&D Systems, F0111).

Western Blot analysis of Carbonic Anhydrase IX/CA9 Antibody on human breast cancer MCF7 cells. Image from verified customer review.

A431 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-Carbonic Anhydrase IX/CA9 Antibody NB100-417 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

An intracellular stain was performed on A431 cells with Carbonic Anhydrase IX/CA9 Antibody NB100-417 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Analysis in 1) HeLa, 2) MDA-MB-231, and 3) A549 whole cell lysates. Specific bands were detected for Carbonic Anhydrase IX/CA9 at a molecular weight of 50 kDa.

Analysis on rat renal cortex. A specific band was detected at a molecular weight of approximately 50 kDa.

Immunocytochemical analysis of CNHCs with antibodies against the RCC marker CAIX. Clusters of CNHCs cytomorphologically classified as uncertain malignant (-UMF) with cytoplasmic positive staining with antibodies against the RCC marker CAIX. Image collected and cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-214) licensed under a CC-BY license.

Analysis of human colon carcinoma, xenografted in mice. IHC-Fr image submitted by a verified customer review.

IHC analysis of a FFPE tissue section of human breast cancer using CAIX antibody at 1:1000 dilution. The primary antibody bound to CAIX antigens in the tissue section was detected using a HRP labeled secondary antibody and DAB reagent. Nuclei of the cells were counterstained with hematoxylin. This CAIX antibody generated an expected cytoplasmic staining of CAIX protein with an intense signal around the cellular membranes in tumor cores. The latter are more likely to be hypoxic in growing tumors which signifies that the observed CAIX staining is specific.

Strong cytoplasmic and membranous CA9 staining in glandular cells of human stomach tissue. Citrate buffer pH 6 antigen retrieval. Antibody at 1:250 dilution and detected with an Alexa Fluor 546 labeled goat-anti-rabbit secondary antibody. Nuclei detected with DAPI. IHC-P image submitted by a verified customer review.

Immunofluorescence of human RCC tumor cryosections using NB100-417 (Panel A). Panel B shows staining with normal rabbit serum.

Colorectal cancer xenograft growth was suppressed by CB-PIC(20 and 50 mg/kg body weight) in female athymic nude mice. Starting three days after SW620 cell inoculation, CB-PIC (20 and 50 mg/kg body weight) was injected in abdomen with 4% Tween 20 as vehicle once daily. Representative examples of immunohistochemical staining for CA IX in tumor section. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/ecam/2013/974313/), licensed under a CC-BY license.

<b>Genetic Strategies Validation. </b>Iron depletion and nitric oxide stress and Iron depletion induce the truncated VDAC1 form accumulation. (D) Total protein extracts were analyzed by immunoblotting using VDACs poly antibody and anti-HIF-1 alpha and -CAIX antibodies. (E) Immunoblotting was used to analyze total protein extracts using antibodies against the three VDAC isoforms. (F) HeLa cells were grown in hypoxia (Hx, 1% O2) conditions and transfected with iscu- or NC-siRNA for 3 days, or grown in normoxia (Nx, 21% O2), some treated with DFO for 16 h. Western Blots analyzed total proteins using VDACs poly antibody and anti-HIF-1 alpha, -CAIX, -ISCU antibodies. Citation: Ferecatu I, Canal F, Fabbri L, Mazure NM, Bouton C, Golinelli-Cohen M-P (2018) Dysfunction in the mitochondrial Fe-S assembly machinery leads to formation of the chemoresistant truncated VDAC1 isoform without HIF-1 alpha activation. PLoS ONE 13(3): e0194782.

The images are representative of immunohistochemistry staining of nuclear staining of proliferation marker Ki67, membrane protein CD 44, and CA IX. The images of scaffold culture (left panel) are compared with cell pellet paraffin section (right panel). The scaffold dissolved in xylene presented as a clear region marked ∗. All scale bars are 100 μm, magnification 40x.

Chemosensitivity of the CTC SCLC lines (IC50, mean ± SD) and immunohistochemistry of sections of tumorospheres. All differences between single cells (SC) and tumorospheres (TS) are statistically significant. Immunohistochemical staining of sections of UHGc5, BHGc16 and BHGc26 CTCs was performed using antibodies directed to CD56, CHGA, CAIX and Ki67, respectively.

Rhebl1 re-introduction restored mitochondrial biogenesis in muscle-specific TAZ knockout mice.a RNA was isolated from gastrocnemius muscle of wild type (WT) and muscle-specific TAZ knockout (mKO) mice intramuscularly transduced with pAAV6-CMV (mKO-Con) or pAAV6-Rhebl1 (mKO-Rhebl1) virus. Levels of Rhebl1 transcripts were analysed by quantitative reverse transcription (qRT-PCR) to confirm Rhebl1 rescue. n = 6 for each condition (WT vs. mKO-Con; *p = 0.0377, mKO-Con vs. mKO-Rhebl1; ****p < 0.0001). b Protein extracted from gastrocnemius of mice in (a) was analysed by immunoblot assays to confirm levels of the indicated proteins. Alpha-tubulin was used as a loading control. Representative data was shown and experiment was performed twice with similar results. c Gastrocnemius muscle of mice in (a) was assessed by immunofluorescent staining for cytochrome c oxidase subunit 2 to visualise mitochondria. Cell nuclei were counterstained with DAPI. Scale bar = 50 μm. Fluorescence was quantified using ImageJ. n = 4 for each condition. (WT vs. mKO-Con; ****p < 0.0001, mKO-Con vs. mKO-Rhebl1; ***p = 0.0002). d RNA isolated from gastrocnemius muscle of mice described in (a) was assessed by qRT-PCR to determine transcript levels of mitochondria-encoded genes. n = 6 for each condition (mt-Atp6; **p = 0.0054 for WT vs. mKO-Con, *p = 0.0378 for mKO-Con vs. mKO-Rhebl1, mt-Co2; ***p = 0.0006 for WT vs. mKO-Con, *p = 0.0185 for mKO-Con vs. mKO-Rhebl1, mt-Cytb; *p = 0.0103 for WT vs. mKO-Con, *p = 0.033 for mKO-Con vs. mKO-Rhebl1, mt-Nd5; **p = 0.0049 for WT vs. mKO-Con). e Genomic DNA isolated from gastrocnemius muscle of mice described in (a) was analysed by quantitative PCR for relative quantification of mitochondrial DNA copy number. n = 6 for each condition (WT vs. mKO-Con; **p = 0.0026, mKO-Con vs. mKO-Rhebl1; *p = 0.039). For (a, c–e), data are shown as means ± SEM. Statistical significance was analysed via one-way ANOVA with Tukey’s multiple comparison test. Source data are provided as a Source data file. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35115527), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Pl

Species Glossary

Reactivity: Hu, Mu, Rt, Pl

Applications:

, , , , , , , , , , , ,

Applications: , , , , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-417AF350
Alexa Fluor 405	NB100-417AF405
Alexa Fluor 488	NB100-417AF488
Alexa Fluor 532	NB100-417AF532
Alexa Fluor 594	NB100-417AF594
Alexa Fluor 647	NB100-417AF647
Alexa Fluor 700	NB100-417AF700
Alexa Fluor 750	NB100-417AF750
Allophycocyanin	NB100-417APC
Allophycocyanin/Cy7	NB100-417APCCY7
Biotin	NB100-417B
DyLight 350	NB100-417UV
DyLight 405	NB100-417V
DyLight 488	NB100-417G
DyLight 550	NB100-417R
DyLight 594	NB100-417DL594
DyLight 650	NB100-417C
DyLight 680	NB100-417FR
DyLight 755	NB100-417IR
FITC	NB100-417F
HRP	NB100-417H
Janelia Fluor 549	NB100-417JF549
Janelia Fluor 646	NB100-417JF646
PE	NB100-417PE
PE/Atto594	NB100-417PEATT594
PE/Cy5.5	NB100-417PECY55
PE/Cy7	NB100-417PECY7
PerCP	NB100-417PCP
mFluor Violet 450 SE	NB100-417MFV450
mFluor Violet 500 SE	NB100-417MFV500
mFluor Violet 610 SE	NB100-417MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-417SS

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NBP1-04676

PGC1 alpha Antibody - BSA Free

(11 Reviews)

(261 Publications)

0.1 ml

NBP1-04676

Mitochondrial content and biogenesis. Immunoblot analysis of PGC1 alpha in MEFs (n = 7; t = 5.764; df = 6). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-12662-3), licensed under a CC-BY license.

HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-PGC1 alpha Antibody NBP1-04676 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Analysis of a FFPE section of mouse prostate using rabbit polyclonal PGC1 alpha Antibody at 1:200 dilution. The antibody generated an expected strong nuclear (punctate in some cells) and a weak cytoplasmic staining in the glandular cells lining of tubule-alveolar gland.

An intracellular stain was performed on A431 cells with PGC1 alpha Antibody NBP1-04676 (blue) and matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

<b>Biological Strategies Validation. </b>Vehicle or RU 486 (5 mg/kg) injected mice were presented with/without corticosterone (10 mg/kg) for 3 days. The expressions of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1 alpha NBP1-04676) and NIX (BNIP3L NBP1-88558) were visualized via western blotting. Loading control is beta-actin. n = 5. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33473105/) licensed under a CC-BY license.

Total protein from human adipose and skeletal muscle tissue, HeLa and A431 cells lines was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/mL PGC1 alpha Antibody (Molecular weight: 91 KDa) in 1% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence

Western Blot of total protein from murine skeletal muscle tissue using PGC1 alpha Antibody. Image from verified customer review.

HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with PGC1 alpha Antibody [NBP1-04676] at a 1:200 dilution overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) [NB100-690] was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Analysis of PGC1 alpha Antibody in mouse prostate using DAB with hematoxylin counterstain.

An intracellular stain was performed on HeLa cells with NBP1-04676AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

An intracellular stain was performed on HepG2 cells with PGC1 alpha Antibody and a matched isotype control. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody.

Western blot shows lysates of A431 human squamous carcinoma parental cell line and PGC1 alpha knockout (KO) A431 cell line. PVDF membrane was probed with 1:1000 of Rabbit Polyclonal PGC1 alpha Antibody (Catalog # NBP1-04676) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog #HAF008). Specific band was detected for PGC1 alpha at approximately 105 kDa (as indicated) in the parental A431 cell line, but is not detectable in the knockout A431 cell line. This experiment was conducted under reducing conditions.

DM1-derived fibroblasts have no changes in mitochondria biogenesis. (A, B) Representative images of immunofluorescence of TOMM20, and PGC1-alpha in DM1 and control fibroblasts (n=3). (C) Medium fluorescence intensity of MitoTracker Red FM in control (n=3) and DM1 cells (n=5) and (D) of Rhodamine 123 in DM1 and control fibroblasts (n=3). (E) mRNA levels of TFAM transcription factor (n=3). (F) mRNA levels of OPA1, MFN1, MFN2, DRP1 and PARKIN in DM1 and control fibroblasts (n≥2).

Feeding the diabetes mellitus (DM) mice with the RSV-enriched diet improved mitochondrial biogenesis in the quadriceps muscle. A) Representative western blots using antibodies against nuclear respiratory factor (NRF-1), ATP synthase, H+ transporting, mitochondrial F1 complex, beta polypeptide (ATP5B), cytochrome c oxidase subunit IV (Cox IV), peroxisome proliferator-activated receptor-gamma coactivator 1-alpha (PGC-1alpha), and GAPDH. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29578301), licensed under a CC-BY licence.

Mitochondrial content and biogenesis. (d) Immunoblot analysis of PGC1alpha in MEFs (n = 7; t = 5.764; df = 6). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28970491), licensed under a CC-BY licence.

Atg7 deficiency in vascular smooth muscle cells (VSMCs) alters mitochondrial quality control and enhances apoptosis. c) Western blot analyses of the expression of TFEB and PGC-1-alpha proteins in aortic VSMCs isolated from Atg7+/+Tagln-Cre+, ApoE-/- and Atg7F/FTagln-Cre+, ApoE-/- mice after 10 weeks of HFD. Cells were incubated either with or without oxidized LDL (200 ug ApoB/mL) for 8 h. beta-Actin was used as the loading control. The graph represents the densitometric analysis of the expression level of TFEB and PGC-1-alpha proteins. The data are expressed as mean +/- SEM of four independent experiments from different primary VSMC cultures. *P 12 in each group. (B) Western blot images using S100 beta , tubulin beta‐3, and beta ‐actin antibodies to total protein lysate in hippocampal area. The statistical results of internal standardized S100 beta / beta ‐tubulin ratio by beta ‐actin levels relative to young adult wild‐type C57BL/6J. White and black bars indicate the wild‐type C57BL/6J and Tet‐mev‐1 mice, respectively. Data are expressed as mean ± SD; *P 12 in each group. (C) Micrographs of immunohistochemical analysis on paraffined hippocampal tissue sections using GFAP and S100 beta antibody. Brown cells indicate GFAP‐stained astrocytes. Scale bar = 100 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27623715), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

At the time of second passage, vessels in successfully growing xenografts of four different tumor types surveyed were of murine origin: A-C Colon; D-F Lung; G-I Pancreatic; J-L Renal Cell Carcinoma. For each tumor type, a representative section of an original patient specimen labeled with anti-huCD31 is shown (A, D, G, J). For each tumor, sections of the first passage xenografted tumor when it was resected are also shown; while no huCD31(+) vessels were identified in the xenografts (B, E, H, K), msCD34(+) vessels were abundant (C, F, I, L). (bars = 50 μ). Image collected and cropped by CiteAb from the following open publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Knockdown of autophagy-related gene 5 (ATG5) inhibits trehalose-induced autophagy in normal human primary airway epithelial cells.Normal human tracheobronchial epithelial cells were transfected with Naito1 chimera RNAi (control siRNA) or ATG5 chimera siRNA (ATG5 siRNA). Twenty-four hours after siRNA transfection, cells were treated with medium or trehalose (TRE, 100 mM) for 48 h. ATG5 protein (A) and LC3 I and LC3 II proteins (B) were examined by Western blot analysis with GAPDH protein used as loading control. The representative Western blot picture was shown from 2 independent experiments with each being performed in triplicate wells. Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.pone.0124524), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Expression of ER stress associated protein and autophagy related protein in the C28I2 chondrocyte infected with Ad-ATG5 or Ad-ATG7. a The ATG5 and ATG7 mRNA level were analysed by RT-PCR in C28I2 cells. b Analysis of ATG5 and ATG7 mRNA level with Q-PCR in C28I2 cells. Ad-ATG5(a), Ad-ATG7(b). c The ATG5 protein expression was analysed by western blotting in the Ad-ATG5 infected C28I2 cells. The level of ATG5 was normalized to beta -actin. d The ATG7 protein expression was analysed by western blotting in the Ad-ATG7 infected C28I2 cells. The level of ATG7 was normalized to beta -actin. e The expression levels of LC3, P62, PERK, p-PERK, Nrf2 proteins after infected with Ad-ATG5 in C28I2 cells for 24 h by western blotting. The levels of related proteins were normalized to beta -actin. f The expression levels of LC3, P62, PERK, p-PERK, Nrf2 proteins after infected with Ad-ATG7 in C28I2 cells for 24 h by western blotting. The levels of related proteins were normalized to beta -actin. *P 100 mitochondria). Data analysis was performed by using one-way ANOVA (∗P 250 cells/chains per sample). c Representative whole-mount IF of adult Plzf-mC/CreER; Z/EG tubules D3 post-TAM (n = 2 mice). Arrowheads: PDX1+ As. Inset shows detail of indicated area. Scale bar, 50 μm. d Representative IF of adult WT testis sections (n = 4 mice). Insets: details of selected areas. EOMES+ GFR alpha 1+ (arrowheads) and EOMES− GFR alpha 1+ cells (bracket) are shown. Tubule stage is indicated. Scale bar, 50 μm. e Representative IF of adult Oct4-GFP testis section (n = 2 mice). Insets: immunostaining within indicated area. EOMES+ GFP− (arrowheads) and EOMES− GFP+ (bracket) spermatogonia are indicated. Asterisks: autofluorescent interstitium. Scale bar, 50 μm. f Representative flow cytometry of fixed and permeabilized testis cells from Oct4-GFP adults (n = 2). PLZF+ cells shown. g Representative IF of adult WT testis sections (n = 4 mice). Insets: immunostaining within indicated area. PDX1+ EOMES+ spermatogonia are indicated (arrowheads). Tubule stage is shown. Asterisk: autofluorescent interstitium. Scale bar, 50 μm. h Representative flow cytometry of fixed and permeabilized WT adult testis cells (n = 4). Mean numbers of EOMES+ and PDX1+ cells in PDX1+ and EOMES+ gates respectively are shown ± s.e.m. i Representative flow cytometry for KI67 in indicated PLZF+ populations of fixed and permeabilized adult WT testis cells (n = 4 mice). j Quantification of flow cytometry from i. Percentage of indicated PLZF+ fractions KI67+ . Horizontal bars: mean values (n = 4 mice). k Representative IF of adult WT testis sections demonstrating PDX1+ Aundiff localisation (arrowheads) within tubules (n = 4 mice). Asterisks: autofluorescent interstitium. Tubule stages are shown. Scale bar, 50 μm. l Quantification of spermatogonial localisation from k. Mean values ± s.e.m. are shown (n = 4 mice, 56–95 tubule sections per mouse). Significance vs. tubule-tubule localisation is indicated. m Representative IF of Id4IRES-GFP adult testis sections (n = 4 mice). Insets show immunostaining within indicated area. Arrowheads: PDX1+ EOMES+ GFP+ spermatogonia. Asterisk: PDX1low EOMES+ GFP+ cell. Tubule stage is shown. Scale bar, 50 μm. n Representative flow cytometry of indicated PLZF+ populations of fixed and permeabilized adult Id4IRES-GFP testis cells (n = 4 mice). Mean numbers of PDX1+ and EOMES+ Aundiff expressing GFP ± s.e.m. o Flow cytometry of Id4IRES-GFP testis cells as in n. PLZF+ fractions are shown. Mean numbers of PLZF+ GFP+ cells positive for EOMES and PDX1 are indicated ± s.e.m. (n = 4 mice). Significance was calculated by two-tailed Student’s t-test (***P < 0.001, ****P < 0.0001) Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/s41467-018-04827-z), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

A431 human skin carcinoma cell line was stained with Rabbit anti-PGC1 alpha Affinity-purified Polyclonal Antibody conjugated to Phycoerythrin (Catalog # NBP1-04676PE, blue histogram) or matched control antibody (Catalog # NBP2-24983, orange histogram).

Reactivity:

Hu, Mu, Rt, Po, Gt, Ha, Sh, Sq, Ca, Eq, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Gt, Ha, Sh, Sq, Ca, Eq, Pm

Applications:

, , , , , , , ,

Applications: , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP1-04676AF350
Alexa Fluor 405	NBP1-04676AF405
Alexa Fluor 488	NBP1-04676AF488
Alexa Fluor 532	NBP1-04676AF532
Alexa Fluor 594	NBP1-04676AF594
Alexa Fluor 647	NBP1-04676AF647
Alexa Fluor 700	NBP1-04676AF700
Alexa Fluor 750	NBP1-04676AF750
Allophycocyanin	NBP1-04676APC
Biotin	NBP1-04676B
DyLight 350	NBP1-04676UV
DyLight 405	NBP1-04676V
DyLight 488	NBP1-04676G
DyLight 550	NBP1-04676R
DyLight 594	NBP1-04676DL594
DyLight 650	NBP1-04676C
DyLight 680	NBP1-04676FR
DyLight 755	NBP1-04676IR
FITC	NBP1-04676F
HRP	NBP1-04676H
Janelia Fluor 549	NBP1-04676JF549
Janelia Fluor 646	NBP1-04676JF646
PE	NBP1-04676PE
PerCP	NBP1-04676PCP
mFluor Violet 450 SE	NBP1-04676MFV450
mFluor Violet 500 SE	NBP1-04676MFV500
mFluor Violet 610 SE	NBP1-04676MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP1-04676SS

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BC100-494

PINK1 Antibody - BSA Free

(13 Reviews)

(247 Publications)

0.1 ml

BC100-494

$PINK1/Parkin-mediated mitophagy. Western Blot of mitochondrial and cytosolic fractions for PINK1, Parkin, and ubiquitin. E Quantitation of protein levels, normalized to VDAC and GAPDH, Ubiquitin: n = 6, PINK1: n = 5, Parkin: n = 8. R, post-reoxygenation. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33980811/) licensed under a CC-BY license.$

Confocal microscopy analysis of the mitophagy initiation in the RPE cells by staining PINK1 and PARKIN. One-year-old WT and dKO mice focusing on the RPE cells in the vicinity of the optic nerve (a,e). PINK1 (b, red) and PARKIN (c, green) were double-stained and the merged image (d) was used to count the colocalized puncta from WT. Similarly, in dKO PINK1 (f, red) and PARKIN (g, green) were double-stained, and the merged image (h) was used to count the colocalized puncta. dKO = NFE2L2/PGC1a double knockout. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32183173/) licensed under a CC-BY license.

Western blot image of PINK1 antibody (BC100-494) in multiple cells lines. Human HeLa (lane 1), Mouse NIH-3T3 (lane 2), L929 (lane 3) and Rat PC12 (lane 4) whole cell protein were separated by SDS-PAGE on a 7.5% polyacrylamide gel. Protein was transferred to PVDF membrane and probed with 2 ug/mL BC100-494 in 1% BSA and detected with an HRP-conjugated anti-rabbit secondary antibody using chemiluminescence. Observed molecular weight ~55 kDa (arrowhead).

Analysis of PINK1 in mouse liver and hypatocytes using PINK1 antibody. Observed molecular weight ~55 kDa. WB image submitted by a verified customer review.

Pathogenic mutants of Parkin are subjected to Ser65 phosphorylation. Phos-tag Western blotting for Parkin and Western blotting for PINK1 were performed using Parkin WT and a series of pathogenic mutants. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep01002) licensed under a CC-BY license.

Alteration of mitochondria and PD associated proteins in SH-SY5Y cells with telomere removal by CRISPR-Cas9. Representative Western blot of PGC-1alpha, NRF1, and PINK1 in SH-SY5Y cells transfected with either gTel or gCont (72 h). Beta-actin served as a loading control. Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/1422-0067/18/10/2093), licensed under a CC-BY license.

35 ug Neuro2A whole cell lysate separated on 8% PAGE and stained for PINK1 (1:1000 in 5% milk powder in TBST; secondary antibody AP at 1:5000). WB image submitted by a verified customer review.

Rabbit heart tissue. IHC-P image submitted by a verified customer review.

HEK293 cells were co-transfected with PINK1 siRNA (#1 or #2) or scrambled siRNA (scrambled) and untagged wild-type (WT) or Ser65Ala (S65A) mutant Parkin as indicated using TransFectin reagent (Bio-Rad). 48 hrs post-transfection, cells were treated with/without 10 uM CCCP for 3 h. 0.25 mg of 1% Triton whole-cell lysate were subjected to immunoprecipitation with GST-Parkin antibody (S966C) covalently coupled to protein G Sepharose and then immunoblotted with anti-phospho-Ser65 antibody in the presence of dephosphorylated peptide. 5% of the IP was immunoblotted with total anti-Parkin antibody. 0.25 mg of whole-cell lysates were immunoprecipitated with anti-PINK1 antibody (S085D) and immunoblotted with anti-PINK1 antibody. Representative of three independent experiments. Image collected and cropped by CiteAb from the following publication (//rsob.royalsocietypublishing.org/cgi/doi/10.1098/rsob.120080) licensed under a CC-BY license.

Characterisation of mitochondrial content and function in PINK1-/- platelets. Cropped immunoblots showing PINK1 expression in 10 uM CCCP (6 hours) treated WT, but not KO platelets by IP (upper panel) with loading control of IP inputs by blotting for PINK1 (lower panel). Uncropped blots for PINK1 are provided in Supplementary Fig. S1b. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-018-32716-4), licensed under a CC-BY license.

Representative TEM picture. The red and green arrows indicate PINK and PARKIN immunogold particles in the dKO RPE samples, respectively (e). ML, melanosome. Scale = 0.2 um. Mitochondria (M). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32183173) licensed under a CC-BY license.

Protein content and oxidation. Gastrocnemius muscle was analyzed by immunoblotting (A) for oxidatively modified protein, STIM1, PARKIN, PINK1, PGC‐1 alpha , VDAC1, and 4HNE (B). Data are means ± SEM. *Significantly different from Tp53inp1+/+.

USP30 restricts basal pexophagy. H). Quantification of Keima-SKL

$The FANCD2 NLS is required for efficient FANCD2 and FANCI monoubiquitination and chromatin association.(A) FA-D2 cells stably expressing LacZ, FANCD2-WT, FANCD2-K561R, FANCD2-∆N57, FANCD2-∆N100 and FANCD2-3N were incubated in the absence and presence of 250 nM MMC for 18 h, and whole-cell lysates were immunoblotted with antibodies to FANCD2, V5, FANCI and alpha -tubulin. The FANCD2 and FANCI L/S ratios are the ratios of monoubiquitinated to nonubiquitinated protein, and were calculated by measuring protein band intensities using ImageJ image processing and analysis software (//rsb.info.nih.gov/ij/). (B and C) FA-D2 cells stably expressing FANCD2-WT, FANCD2-∆N57, FANCD2-∆N100 and FANCD2-3N were treated as above and cell pellets were fractionated into soluble (S) and chromatin-associated (C) fractions. Fractions were immunoblotted with antibodies against V5, FANCI, alpha -tubulin and H2A. W, unfractionated whole cell extract. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/24278431), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Immunoblot analyses of protein expression for Nrf2 and antioxidant enzymes.A. Analysis of nuclear Nrf2 in young and old mice subjected to EES. In sedentary mice, Nrf2 protein levels were decreased significantly in young versus old mice. EES exacerbated the decrease of nuclear Nrf2 in old mice. Blots/values represent n = 4–6 from each group. *p10 embryos/sample). The blots were probed with antibodies against Lc3 and Actin as a loading control. Western blots were repeated at least three times with independent extracts. (C) Quantification of Lc3-II fold changes in WT and mutant embryos in absence or presence of Baf A1. Western blot band intensities were quantified by Lab Image. Data is combined from three independent experiments. (D) Representative confocal micrographs of GFP-Lc3 puncta present in the tail fin of optn+/+, optn delta 5n/ delta 5n, p62+/+ and p62 delta 37n/ delta 37n at 4 dpf. Scale bars, 10 μm. (E). Quantification of the number of GFP-Lc3 puncta in optn+/+, optn delta 5n/ delta 5n, p62+/+ and p62 delta 37n/ delta 37n larvae with and without Baf A1 treatment. Each larva was imaged at a pre-defined region of the tail fin (as indicated by the red boxed area in Fig3 A) (≥11 larvae/group). Results are accumulated from two independent experiments. ns, non-significant, *p 12 months) were injected i.v. with siScr or siMST. Twenty‐four hours after siRNA injection, a 70% PH was performed and remnant/regenerating liver tissue was harvested 40 h later. p.i. of the livers was calculated by IHC staining of Ki67‐positive hepatocytes and livers were classified as (−) non‐regenerating or (+) regenerating based on the percentage of positive cells. Non‐surviving animals were sacrificed before the 40‐h end time point. “n” indicates the number of animals per group.Ki67 immunostaining of the regenerating aged mice livers, 40 h post‐PH. Sections were scanned using 3DHistech Pannoramic MIDI Scanner and quantitated with Quant Center 2.0 software. p.i. values are provided on the image.Representative photomicrograph of an aged liver treated with siMST and stained with H&E and by IHC for CK19 and Ki67. Arrows indicate ductal regions with no signs of reaction or oval cell expansion.Quantitative RT–PCR analysis of RNA isolated from mouse liver for Foxm1B, Birc5 and Ccnb1 at 0 and 40 h post‐PH. Log2 fold change was calculated using non‐transfected/non‐resected mouse liver as a control. Representative results from a single experiment with n = 6 animals per group are shown. Paired, two‐tailed Student's t‐test was used to calculate the significant change in signals between liver tissues at the time of resection (0 h) compared to 40 h post‐PH.Percentage of Ki67‐positive hepatocytes in aged and aged + siMST 40 h post‐resection. Representative results from a single experiment with n = 6 independent aged animals for group are shown. Two‐tailed Student's t‐test was used to calculate the significance of percentage of positive hepatocytes of siMST‐treated animals compared to non‐treated controls.Endpoint liver weight was taken of the remnant lobe and expressed as a percentage to total body weight. Representative results from a single experiment with n = 6 independent animals for group expect for control group where n = 6 are shown. Estimated liver‐to‐body weight ratio before PH in mice = 3.85 ± 0.05 (SD). Unpaired, two‐tailed Student's t‐test was used to calculate the significance of each aged group in comparison with the control young group.Hepatocyte geometric diameter was determined (see Materials and Methods). Representative results from a single experiment with n = 3 independent animals per group. Unpaired, two‐tailed Student's t‐test was used to calculate the significant change in hepatocyte size in liver tissue with and without siMST 40 h post‐PH.Data information: Bars represent mean ± SD. Scale bars, 50 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27940445), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, Rb

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	BC100-494AF350
Alexa Fluor 405	BC100-494AF405
Alexa Fluor 488	BC100-494AF488
Alexa Fluor 532	BC100-494AF532
Alexa Fluor 594	BC100-494AF594
Alexa Fluor 647	BC100-494AF647
Alexa Fluor 700	BC100-494AF700
Alexa Fluor 750	BC100-494AF750
Allophycocyanin	BC100-494APC
Biotin	BC100-494B
DyLight 350	BC100-494UV
DyLight 405	BC100-494V
DyLight 488	BC100-494G
DyLight 550	BC100-494R
DyLight 594	BC100-494DL594
DyLight 650	BC100-494C
DyLight 680	BC100-494FR
DyLight 755	BC100-494IR
FITC	BC100-494F
HRP	BC100-494H
Janelia Fluor 549	BC100-494JF549
Janelia Fluor 646	BC100-494JF646
PE	BC100-494PE
PerCP	BC100-494PCP
mFluor Violet 450 SE	BC100-494MFV450
mFluor Violet 500 SE	BC100-494MFV500
mFluor Violet 610 SE	BC100-494MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	BC100-494SS

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NB400-104

SR-BI Antibody - BSA Free

(5 Reviews)

(227 Publications)

0.1 ml

NB400-104

Immunocytochemistry/Immunofluorescence: SR-BI Antibody [NB400-104] - SR-BI antibody was tested in human fibroblast samples fixed in 4% PFA and permeabilized in PBS (0.2% Tween). Primary incubation overnight at 4C using a 1:100 dilution in PBS (0.1% Tween) with 1% BSA. Secondary antibody is anti-rabbit conjugate to Alexa Fluor 488. SR-BI is shown in green and nucleus in blue (Hoescht 33342 stain).

Flow Cytometry: SR-BI Antibody [NB400-104] - An intracellular stain was performed on HeLa cells with SR-BI antibody NB400-104AF488 (blue) and a matched isotype control NBP2-24893AF488 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

Western Blot: SR-BI Antibody [NB400-104] - Detection of SR-BI in rat H4IIE total cell lysates and plasma membrane proteins. Photo courtesy of product review by verified customer.

Immunohistochemistry: SR-BI Antibody [NB400-104] - Immunolocalization of SR-BI in adult mink testis using NB400-104. SR-BI labeling is visible at the surface and along the outline of the large vacuole. Photo courtesy of R.M. Pelletier, University of Montreal.

Immunohistochemistry-Paraffin: SR-BI Antibody [NB400-104] - FFPE tissue section of mouse liver using SR-BI antibody (Lot 8310) at 1:300 dilution with HRP-DAB detection and hematoxylin counterstaining. The antibody generated a specific membrane signal of SR-BI protein in the murine hepatocytes.

<b>Genetic Strategies Validation. </b>Knockdown Validated: SR-BI Antibody [NB400-104] - HCV replication was increased by CD36. Silence of CD36 decreased HCV infection and replication (n = 3). Huh7.5 cells were transfected with 0.2 u g CD36-HA plasmid or with 150 pmol siRNA for CD36 (B) for 48 hrs, and then were washed and infected with HCV (150 IU/cell) for 2 hrs, followed by washing and continuously incubating. Intracellular HCV RNA and proteins were detected in 96 hrs, and the cytotoxicity of siRNAs was measured in 96-wells plate with a MTT assay in B. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep21808) licensed under a CC-BY license.

Immunohistochemistry-Paraffin: SR-BI Antibody [NB400-104] - Staining of human adrenal cortex.

Immunocytochemistry/Immunofluorescence: SR-BI Antibody [NB400-104] - HeLa cells were fixed and permeabilized for 10 minutes using -20C MeOH. The cells were incubated with anti-SR-BI at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry-Paraffin: SR-BI Antibody [NB400-104] - FFPE tissue section of mouse liver using SR-BI antibody (Lot R-4) at 1:300 dilution with HRP-DAB detection and hematoxylin counterstaining. The antibody generated mainly a membranous signal of SR-BI protein in the murine hepatocytes.

Flow Cytometry: SR-BI Antibody [NB400-104] - Analysis of Huh7 and HepG2 cells using SR-B1 antibody NB400-104. Courtesy of Bruno Sainz, Jr., PhD, University of Illinois at Chicago.

Immunofluorescence assay on GIM biopsies of HCV-transplanted patients.Immunofluorescence assay on GIM polyp biopsies of HCV-transplanted patients, using antibodies against HCV receptors: CD81, SR-B1, Claudin-1, Occludin (Original magnification X400). (A, E) CD81 in colon (patient 26); (B, F) Claudin-1 in colon (patient 26); (C, G) Occludin in colon (C patient 28, G patient 26); (D) SR-B1 in antrum (patient 28); (H) SR-B1 in colon (patient 26). All the sections were clearly positive for the analyzed HCV receptors before (A-D) and after transplantation (E-H). Negative controls (I-N) were performed on GIM biopsies of HCV positive patients by omitting the primary antibodies, and by using polyclonal FITC-conjugated Donkey anti-mouse (I) and anti-rabbit (L, M, N) as secondary antibodies. (I CD81 in antrum; L Claudin-1 in polyp colon; M Occludin in antrum; N SR-B1 in polyp colon. Nuclei were counterstained with DAPI (blue). Scale bar 50 um. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0181683), licensed under a CC-BY licence.

HCV replication was increased by CD36. (B) Silence of CD36 decreased HCV infection and replication (n = 3). Huh7.5 cells were transfected with 0.2 ug CD36-HA plasmid (A) or with 150 pmol siRNA for CD36 (B) for 48 hrs, and then were washed and infected with HCV (150 IU/cell) for 2 hrs, followed by washing and continuously incubating. Intracellular HCV RNA and proteins were detected in 96 hrs, and the cytotoxicity of siRNAs was measured in 96-wells plate with a MTT assay in B. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep21808), licensed under a CC-BY licence.

ALDH2 rs671 modulates HDL-C levels in mice and human liver through increasing poly(ADP-ribosyl)ation of LXRalpha due to attenuated ALDH2/PARP1 interaction. (C) Western blotting analysis of ABCA1, ALDH2, LXRalpha, and SR-B1 expression in mouse liver tissue. WT, n = 3; rs671, n = 3. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35393951), licensed under a CC-BY licence.

ZA treatment increases ABCA1 and apoA-1 expression.(a) Western blot analysis of ABCA1, ABCG1 and SR-BI expression in untreated (ZA-) and ZA-treated (ZA+) THP-1 cells, U937 cells, CD14+ cells and corresponding DC subsets (DCTHP1, DCU937, DC). ABCA1 was already detectable in DCU937 and DC and its expression was upregulated by ZA. beta-tubulin was employed to check the equal protein loading per lane. The results are obtained from one representative experiment of three experiments. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms15663), licensed under a CC-BY licence.

Differentiation and characteristics of iHLCs. (d) Western blot analysis of protein expression in HCV-infected iHLCs compared to Huh7.5 cells of different host factors crucial for HCV infection. Tubulin served as loading control. Full-length blots are presented in Supplementary Figure 1. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29497123), licensed under a CC-BY licence.

Expression of cellular HCV receptors at the mRNA and protein levels.(a) mRNA copy numbers of CD81, occluding (OCLN), claudin-1 (CLDN1) and SR-BI were quantified in OASF, RASF and Huh-7.5 cells, normalized to the reference gene GAPDH. (b) Detection of CD81 on Huh-7.5 cells, OASF, and RASF by FACS analysis. (c-d) Detection of OCLN, CLDN1 and SR-BI in OASF and RASF by immunoblot analysis. Huh-7.5 cells were used as positive control and Hep-56.1D as negative control. In both the real-time PCR and the FACS analysis experiments the number of biological replicates was 3 and the number of experimental replicates was 2 (total n = 6). In immunoblotting detection, the number of biological replicates was 2 and the number of experimental replicates was 2 (n = 4). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26643193), licensed under a CC-BY licence.

miR-27a/b inhibit PINK1 accumulation upon mitochondrial damage. a, b Overexpression of miR-27a/b inhibited PINK1 accumulation upon mitochondrial damage. 48 h post-transfection, HeLa cells were treated with 10 μM CCCP or combination of 10 μM oligomycin and 4 μM antimycin as indicated for 2 h (n = 4, two-way ANOVA). c, d Inhibition of endogenous miR-27a/b increased PINK1 accumulation upon CCCP treatment. (n = 4, two-way ANOVA). PINK1 levels were normalized to corresponding GAPDH level and quantified as a percentage of control. Values are mean ± SEM (n.s. = non-significant, *p < 0.05, **p < 0.01, ***p < 0.001) Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27456084), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

A beta induces the expression of cell cycle regulatory proteins. (A) SK-N-MC cells were exposed to A beta (5 μM) for 24 h. The mRNA expression levels of CDK5, P35 and P39 were analyzed by real-time PCR. The mRNA expression level was normalized by beta -actin mRNA expression level. Data represent the mean ± SE. n = 4. (B)hif1 alpha specific- and non-targeting (NT) siRNA were transfected to the cells for 24 h prior to A beta treatment. Cyclin D1, CDK4, cyclin E, CDK2, HIF1 alpha and beta -actin was detected by western blot. n = 3. (C–F) Cells were pretreated with trehalose (10 μM), rapamycin (10 nM), PF4708671 (10 μM) and cycloheximide (4 μM) for 30 min prior to A beta treatment for 24 h. Cyclin D1, CDK4, cyclin E, CDK2 and beta -actin were detected by western blot. n = 3–6. (G) Mouse hippocampal neurons were transfected with hif1 alpha specific- and NT siRNAs for 24 h prior to A beta treatment for 24 h. Samples were blotted with Cyclin D1, CDK4, cyclin E, CDK2 and beta -actin specific antibodies. n = 3–6. (H) Mouse hippocampal neurons were pretreated with trehalose (10 μM) for 30 min and incubated with A beta for 24 h. cyclin D1, CDK4, cyclin E, CDK2, HIF1 alpha and beta -actin were analyzed by western blot. n = 3–6. Data are presented as a mean ± SE. *p < 0.05 vs. control, #p < 0.05 vs. A beta treatment. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28790888), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Yap deficiency suppresses cell proliferation in vivo and in vitro. (A-F) The Ki67 positive ratio of lens epithelial cells decreased in Yap-deficient mice at different stages (arrowheads indicate Ki67 positive cells). (G) The relative number of Ki67 positive lens epithelial cells (number of Ki67 positive lens epithelial cells / lens epithelium area). The data are shown as mean ± S.E.M. (Student’s t-test, *P<0.05, **P<0.01, n=10). (H-I) Knockdown efficiency of Yap in alpha TN4 cell using siRNA. (J-K) Cell viability and growth assay revealed that proliferation was downregulated in Yap knockdown alpha TN4 cells. The data are shown as mean ± S.E.M. (Two-way RM ANOVA, **P<0.01, n=5). Scale bars: 50 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31011480), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Knockdown of CIP2A inhibited cell proliferation and DNA synthesis of HPV-16E7-expressing cells(A) Western blot analysis of protein level of 16E7 and CIP2A in RPE1-16E7 cells and (B) with CIP2A siRNA for 48 hr. (C) CCK8 assay of cell proliferation of RPE1-16E7 cells with CIP2A siRNA. (D) Flow cytometry of cells with CIP2A siRNA and labeled with BrdU for 2 hr, then stained with PI and BrdU; and (E), Quantification. Babe, vector control. **, P < 0.01. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/25650660), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

ATO interferes with efficient HAdV replication center formation and reorganization of PML‐NBs. H1299 cells were infected with HAdV‐C5 wt at a multiplicity of 20 FFU per cell, treated with the depicted concentrations of ATO at 2 h p.i., fixed 48 h p.i. with 4% PFA and double labeled with mAb B6‐8 ( alpha ‐E2A) and pAb NB100‐59787 ( alpha ‐PML). Primary antibodies were detected using Alexa488 (PML, green) and Alexa647 (E2A, red) conjugated secondary antibodies. A) Number of PML‐NBs per cell in uninfected cells was determined using Volocity for at least n = 461 cells from two independent biological replicates. Statistically significant differences were determined using a one‐way ANOVA and Dunnet's T3 test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001. B) The proportion of infected cells showing track‐like redistribution of PML‐NBs was determined by counting for at least n = 212 cells and normalization to untreated infected cells. C) The proportion of infected cells showing formation of HAdV replication centers marked by the viral protein E2A (lower plot) was determined by counting for at least n = 212 cells and normalization to untreated infected cells. D) Cells showing either viral replication centers with PML track‐like structures, replication centers without PML track‐like structures, no replication centers but PML track‐like structures or no replication centers, and no PML track‐like structures were counted for at least n = 214 (virus infected cells due to E2A signal detected: either untreated/0 µm or treated with ATO/1 or 2 µm) and represented in pie charts. Statistically significant differences were determined using a one‐way ANOVA and Dunnet's T3 test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001. E) H1299 cells were infected with HAdV‐C5 wt at a multiplicity of 20 FFU per cell, and treated with 0 or 2 µm of ATO at 2 h p.i. After 48 h, the cells were fixed with 4% PFA and stained using pAb NB100‐59787 ( alpha ‐PML) and mAb 6A‐11 ( alpha ‐E4orf3). Primary antibodies were detected using Alexa488 (PML, green) and Alexa647 (E4orf3, red) coupled secondary antibodies. Representative staining patterns for at least 30 uninfected cells treated with 0 or 2 µm ATO are shown in panels (a)–(d) and (m)–(p), infected cells treated with 0 or 2 µm ATO are shown in panels (e)–(l) and (q)–(x). Overlays of single fluorescence pictures (merge) are shown in panels (d), (h), (l), (p), (t), and (x). Data corresponds to two independent biological replicates performed and counted by different operators to avoid operator bias. Scale bar represents 10 µm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32328411), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$ABCC4 protein expression level in CRC cell lines. Western blot performed in standard reducing SDS PAGE conditions using goat anti ABCC4 (#PA5 18315, Thermo Scientific) and rabbit anti ABCG2 (#ORB 155559 Biorbyt). (A) Protein expression level of ABCC4 and ABCG2 in HT-29 stably overexpressing transcription factor Snail (HT29/Snail) and control HT-29. ABCC4 level in the membrane fraction (obtained by biotynylation using EZ-Link Sulfo -NHS-Biotin Thermo Scientific kit) of HT-29 control cells and HT-29 Snail n = 3. (B) ABCC4 protein expression level in CRC cells in different states of EMT: CCD841CoN (most epithelial), CaCo-2 (moderate EMT), and Colo-320 (most mesenchymal) n = 3. (C) ABCC4 protein abundance in Extracellular Vesicles (EVs) released from HT-29 control cells and two HT-29 stably overexpressing transcription factor Snail clones (HT-29/Snail and HT-29/Snail17), n = 2. (D) Intracellular cAMP level measurement. Accumulation of cAMP in HT29 cells was measured using a cAMP competitive kit (#581001 Cayman Chemical). Cells were incubated for 24 h with MK571 20 µM, or untreated ones were assayed according to the manufacturer’s protocol. Calculation were conducted using the Cayman data sheet. cAMP concentration of HT29 was set as 100%. T-test performed, n = 5; * p < 0.05; ** p < 0.005; *** p < 0.001. NS—not statistically significant. (E) PKA phosphorylation profile analysis. HT29 Snail cells were seeded on a 6-well plate. Then, 24 h after, full growth medium was changed into starving (FBS free) medium for 24 h. Next, 20uM of MK571 was added to cells for 60, 30, 5, and 1 min. Cells without the starving procedure were used as a positive control, and negative control cells were not treated with MK571. Phosphorylation profile analysis was performed using phospho-(ser/thr) PKA Substrate Antibody #9621 (Cell Signaling Technology). Significant time- (exposure) related impact on the phosphorylation profile was observed for 42 kDa and 95–100 kDa proteins in HT29 Snail cells compared to no time-related changes in control cells, n = 3. (F) HT-29/Snail PKA phosphorylation profile analyzed with densitometry; statistical significance estimated using T-test. * p < 0.05; ** p < 0.005; *** p < 0.001. NS—not statistically significant. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/33261018), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Reactivity:

Hu, Mu, Rt, Po, ChHa, Ha, Md, Pm, Rb, SyHa

Species Glossary

Reactivity: Hu, Mu, Rt, Po, ChHa, Ha, Md, Pm, Rb, SyHa

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB400-104AF350
Alexa Fluor 405	NB400-104AF405
Alexa Fluor 488	NB400-104AF488
Alexa Fluor 532	NB400-104AF532
Alexa Fluor 594	NB400-104AF594
Alexa Fluor 647	NB400-104AF647
Alexa Fluor 700	NB400-104AF700
Alexa Fluor 750	NB400-104AF750
Allophycocyanin	NB400-104APC
Biotin	NB400-104B
DyLight 350	NB400-104UV
DyLight 405	NB400-104V
DyLight 488	NB400-104G
DyLight 550	NB400-104R
DyLight 594	NB400-104DL594
DyLight 650	NB400-104C
DyLight 680	NB400-104FR
DyLight 755	NB400-104IR
FITC	NB400-104F
HRP	NB400-104H
Janelia Fluor 549	NB400-104JF549
Janelia Fluor 646	NB400-104JF646
PE	NB400-104PE
PerCP	NB400-104PCP
mFluor Violet 450 SE	NB400-104MFV450
mFluor Violet 500 SE	NB400-104MFV500
mFluor Violet 610 SE	NB400-104MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB400-104SS

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At	=	A. thaliana	SyHa	=	Golden Syrian Hamster
All	=	All Species	Ha	=	Hamster
An	=	Animal	Hu	=	Human
ArHa	=	Armenian Hamster	I	=	Insect
Av	=	Avian	Ll	=	Llama
Bb	=	Baboon	Ma	=	Mammal
Ba	=	Bacteria	Mk	=	Monkey
Bv	=	Bovine	Mu	=	Mouse
Ca	=	Canine	Multi	=	Multi-species
Ce	=	C. Elegans	NA	=	Non-species specific
Ch	=	Chicken	Or	=	Orangutan
ChHa	=	Chinese Hamster	Pl	=	Plant
Cm	=	Cynomolgus Monkey	Pm	=	Primate
Do	=	Donkey	Po	=	Porcine
Dr	=	Drosophilia	Rb	=	Rabbit
Ec	=	E. Coli	Rt	=	Rat
Eq	=	Equine	RM	=	Rhesus Macaque
Fe	=	Feline	Sh	=	Sheep
Ft	=	Ferret	Vb	=	Vertebrate
Fi	=	Fish	Vi	=	Virus
Fu	=	Fungi	Xp	=	Xenopus
Ge	=	Gerbil	Ye	=	Yeast
GP	=	Guinea Pig	Ze	=	Zebrafish
Gt	=	Goat

Category

Protein/Target

Applications

Reactivity

Citation

Antibody Validation

Reviews

Host

Available Conjugates

Format

Marker

Modification

Clonality

Gene Symbol

Images Type

Isotype

Preservative

Research Areas

Has Sample Size

Promotional Item

Species Glossary

LC3B Antibody - BSA Free

HIF-2 alpha/EPAS1 Antibody - BSA Free

HIF-1 alpha Antibody - BSA Free

ABCA1 Antibody - BSA Free

LC3B Antibody - BSA Free

AIF-1/Iba1 Antibody

IRE1 alpha [p Ser724] Antibody - BSA Free

HIF-1 alpha Antibody

Carbonic Anhydrase IX/CA9 Antibody - BSA Free

PGC1 alpha Antibody - BSA Free

PINK1 Antibody - BSA Free

SR-BI Antibody - BSA Free