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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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NB100-304

53BP1 Antibody - BSA Free

(14 Reviews)

(712 Publications)

0.1 mg

NB100-304

Irradiated cochlear spiral ganglion cells, mouse, frozen section of fixed material. Image from verified customer review.

<b>Genetic Strategies Validation. </b>53BP1 was detected in immersion fixed HeLa cells (left) but was not detected in 53BP1 knockout Hela cells (right) using Rabbit Anti-human 53BP1 polyclonal antibody (Catalog #NB100-304) at 0.3 ug/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei.

Total protein from HeLa, A431, Neuro2A, and PC12 was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 1.0 ug/mL in 5% block buffer and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. The observed molecular weight for these samples are ~250 kDa and the theoretical molecular weight is 214 kDa.

Human breast tumors stained with 53BP1 antibody. Image from verified customer review.

Detection of Human and Mouse 53BP1 by IHC. Sample: FFPE sections of human ovarian carcinoma (left) and mouse teratoma (right). Antibody: 53BP1 Antibody (Catalog #NB100-304) used at a dilution of 1:1000 (1ug/mL). Detection: DAB.

Human medulloblastoma (DAOY) and mouse astrocyte (C8D1A) cell lines were exposed for 48 hours to DMSO or 1ug/mL of the DNA damaging agent EPE. Cells were immunostained for 53BP1 (green). The nuclei were counterstained with DAPI (blue). Image from verified customer review.

An intracellular stain was performed on RH-30 cells with 53BP1 Antibody (Catalog #NB100-304AF647) (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 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Immunoprecipitation analysis lysates from HCC44 cells in 1% NP40. Image from verified customer review.

<b>Biological Strategies Validation. </b>Effect of RK-33 on radiation-induced DNA damage. A Immunofluorescence images showing 53BP1 and gamma-H2AX foci in A549 cells after 2-Gy radiation and A549 cells pre-treated with 6 uM RK-33, 12 h before radiation. Overlap of 53BP1 and gamma-H2AX is seen in the merged picture of the co-immunofluorescence staining. Scale bar is 2 um. Image collected and cropped by CiteAb from the following publication (//embomolmed.embopress.org/cgi/doi/10.15252/emmm.201404368) licensed under a CC-BY license.

Lamin A/C-53BP1 interaction is regulated in a DNA damage-dependent manner. U2OS/GFP-lamin A cells were pretreated with 10 um ATMi f for 1 h before exposure to IR (10 Gy, 1 h recovery). Cell extracts were then subjected to immunoprecipitation using GFP-Trap beads, and bound complexes were then analyzed by immunoblotting using 53BP1 and GFP antibodies. WCE represents 1% input. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1111/acel.12258), 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 53BP1 Antibody conjugated to DyLight 550 (NB100-304R) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

An intracellular stain was performed on Ntera2 cells with 53BP1 Antibody NB100-304 (blue) and a 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).

Whole cell lysate from U2OS or 293T cells. Bands indicate an observed molecular weight of ~220 kDa and the theoretical molecular weight is 214 kDa.

<b>Biological Strategies Validation. </b>Upper Panel: 53BP1 foci in proliferating MEFs. Lower Panel: 53BP1 foci in proliferating MEFs exposed to 10 Gy of IR.

Embryonic Fibroblast cells pre-extraction for 5 mins with CSK buffer. Fixed with 4% PFA and 75% Ethanol. Primary Antibody at 1:1000. Secondary Antibody at 1:1000. Image from verified customer review.

<b>Biological Strategies Validation. </b>Upper Panel: Control untreated cells. Lower Panel: Cells exposed to irradiation (10 Gy) and probed for 53BP1 foci. Cells were grown on coverslips, fixed with 4% paraformaldehyde, methanol permeabilized, blocked for 1 h, RT. Incubated with primary antibody (1:200) overnight, washed 3x with PBS, probed with tubulin (Alexa Fluor 594) antibody for 2 h, RT. Washed 3x with PBS, mounted on slides using prolong gold, imaged using Nikon confocal microscope (100x oil). Image from verified customer review.

<b>Genetic Strategies Validation. </b>MUS81 inhibition in BRCA2-deficient cells causes accumulation of 53BP1 nuclear bodies and G1 arrest. H1299 cells carrying a DOX-inducible BRCA2 shRNA were transfected with control or MUS81 siRNAs. Representative images of cells processed 72 h later for immunostaining with 53BP1 Antibody (green) and anti-cyclin A (red) antibodies. DNA was counterstained with DAPI. Scale bar, 10 um. Image collected and cropped by CiteAb from the following publication (//www.nature.com/doifinder/10.1038/ncomms15983), licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>Pot1a prevents DDR in HSCs. Telomeric DDR in 8 week-old LSK cells upon Pot1a knockdown. Immunocytochemical staining of TRF1 (green). Foci co-stained with TRF1 and 53BP1 were identified as TIFs. Nuclei were stained with TOTO3 (blue). Scale bar, 2 um. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-017-00935-4), licensed under a CC-BY license.

Lamin A/C-53BP1 interaction is regulated in a DNA damage-dependent manner. U2OS cells were transfected with siCTRL or siLMNA and subjected to laser micro-irradiation, fixed 15 min later and then processed for immunofluorescence with gamma-H2AX and 53BP1 antibodies. Scale bar, 10 um. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1111/acel.12258), licensed under a CC-BY license.

Ntera2 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-53BP1 Antibody NB100-304 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. 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 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

NIH3T3 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-53BP1 Antibody NB100-304 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. 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 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Staining of 53BP1 in human colon cancer using DAB with hematoxylin counterstain.

An intracellular stain was performed on A431 cells with 53BP1 Antibody (Catalog #NB100-304AF647) (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.

For 53BP1 staining, samples were incubated for 1 hour with 1:200 53BP1 primary antibody in PBS/BSA 1%, at room temperature. Green: 53BP Blue: DAPI. Image from verified customer review.

DYNLL1 is required for 53BP1-dependent p53 responses to Nutlin-3. a Immunoblot analysis of the MCF-7 cell lines used in (a) with anti-53BP1 or anti-RPA32 antibodies prior to N3 treatment. b Quantification of n = 3 independent experiments represented in (a), each performed in triplicate. Mean ± SD. c Indicated parental or stably complemented 53BP1-/- MCF-7 cell lines were incubated in the presence (11 days) or absence (7 days) of Nutlin-3 (4 µM)

Depletion of PNUTS and NPAT induces a global DDR.(A) Induction of telomere dysfunction–induced foci (TIF) upon candidate knockdown in HeLa cells from Fig 5. Depletion of TRF2 was used as a positive control. 53BP1 was detected by indirect IF and telomeres by FISH with a Cy3-[CCCTAA]3 probe. (B) Frequency distribution of the number of TIFs per cell from (A). n indicates the number of cells analyzed for each condition and red lines mark the mean.

HPV+ HNSCCs have decreased expression of NHEJ and HR proteins including DNA-Pk and BRCA2Cells were treated with mock or 4 Gy radiation, harvested and lysed at 15 minutes post-treatment, and analyzed by western blot for relative expression of indicated proteins. beta -actin was used as a loading control. Shown is a representative blot from 2 independent experiments.

LNCaP cells in culture were treated for 24 hrs with DMSO or ENZ, then irradiated with 0 or 1 Gy.Cell were fixed and immunostained for 53BP1 0.5, 1.5 and 24 hrs after irradiation. Immunofluorescent images were obtained by confocal microscopy. White bar = 20 microns. A) DMSO alone, B) ENZ alone, C) 1 Gy, D) ENZ + 1 Gy. White bar = 20 microns.

Representative immunohistochemistry staining for 53BP1 expression in pancreatic adenocarcinoma. (A) is high intensity of 53BP1 expression and (B) is low intensity of 53BP1 expression.

Absence of Roma Leads to Activation of the DNA Damage Response and Genomic Instability(A) Representative immunofluorescence analysis of RPA2-pT21, gammaH2AX, and 53BP1 foci in tissue sections from WT and Roma-/- glands at 10dL. The white arrow indicates nucleolar gammaH2AX. Scale bars, 10 um. The higher magnification panels on the right have been chosen to highlight the foci and are not zoomed images of the left panels. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27149840), licensed under a CC-BY licence.

Persistent DNA damage exists in ALT cells. (A) IF-FISH showing DNA damage response (53BP1) in ALT (VA13, U2OS, SAOS2, SKLU-1), telomerase positive (MCF7, A549), and human normal fibroblast cells (MRC5, BJ). Scale bar: 10 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31115790), licensed under a CC-BY licence.

Persistence of repair foci at suboptimal pHe. (A) Left panel shows immunofluorescence detection of gammaH2AFX and 53BP1 following 24 h recovery from bleomycin treatment. Control, sample before damage induction. Colocalization is shown in white; PCC, Pearson correlation coefficient score. Right panel corresponding to gammaH2AFX foci/cell at 24 h and 48 h following recovery. Only foci larger than 10 pixels were counted. Experiments were performed in triplicate. Values are mean +/- SEM. *P 3 (c) gamma-H2AX or d 53BP1 foci per nucleus. n = 90 cells from three independent experiments. Mean +/- SEM; two-way ANOVA (gamma-H2AX: F = 35.77, DFn = 4, DFd = 20, p 10 foci was called positive and counted). DAPI was used for nuclear staining. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28489060), licensed under a CC-BY licence.

USP15 regulates homologous recombination (HR) and breast cancer response to poly-(ADP-ribose) polymerase (PARP) inhibitor. c) Non-homologous end-joining (NHEJ) efficiency in USP15-depleted cells were determined using the EJ5-GFP reporter assay, as described in the Methods section. CtIP or 53BP1 small interfering RNAs (siRNAs) were used as a positive control, respectively, in b, c. Data are shown as average of three independent experiments, presented as mean +/- SD, two-tailed Student's t test, *P < 0.01. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30874560), licensed under a CC-BY licence.

MRE11 poly(T)11/12 sequence variant has no apparent functional consequence on DNA damage detection and repair. (D) DNA repair foci was observed by immunofluorescence after 1.5 Gy IR followed by 1 h and 4 h recovery. Irradiation resulted in the formation of both MRE11 and 53BP1 foci in Daudi and Raji cells, whereas Jurkat failed to form MRE11 foci. Nuclei were counterstained with DAPI. Merged figure presents co-localization of MRE11 and 53BP1 foci visualized in yellow. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/21637783), licensed under a CC-BY licence.

Phenotypic consequences of R325Q mutation. (D) DNA damage is increased in patients' B-LCLs as compared to control, with an accumulation of 53BP1 foci. Left: immunostainings of 53BP1 imaged by confocal microscopy, Right: quantification of 53BP1 foci number. Graph represents the average count obtained from two independent experiments (Mean +/- SEM). Total number of cells in control = 508; P2_E71K n = 315; P_R325Q n = 452; Sidak's multiple comparisons test, ****p < 0.0001. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31787977), licensed under a CC-BY licence.

Bystander signaling evokes DNA damage foci formation in urothelial cancer and normal urothelial cells. (A) Schematic representation of experimental conditions including non-irradiated, uniformly-irradiated and shielding protocols where 50% of the slide was exposed and 50% shielded with MCP alloy. Cells were fixed and processed for 53BP1-immunofluorescence, one hour following irradiation. Micrographs of urothelial cancer T24 cells labelled with anti-53BP1 (green) to label nuclear DNA double stand break foci and DAPI to counterstain nuclei are shown for each experimental condition. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29228614), licensed under a CC-BY licence.

Cry2-/- cells accumulate damaged DNA.(A) Representative early passage (P3-4) primary wildtype (WT), Cry1-/-, Cry2-/-, and Cry1-/-;Cry2-/- adult ear fibroblasts stained with anti-53BP1 antibody (green) and DAPI (blue). Insets show enlarged view of indicated cells. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25756610), licensed under a CC-BY licence.

Chromatibody-driven DDR alteration. (B) RNF8-dependent H2A di-ubiquitylation. HT1080 cells were transfected with the indicated constructs for 24 h. Whole-cell extracts were analyzed with an anti-ubiquitinylated H2A antibody (UbH2A). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27206857), licensed under a CC-BY licence.

The susceptibility of human LECs to low-dose ionizing radiation. The human lens cell line FHL124 was exposed to low-dose IR up to 2.28 Gy. Exposed cells were then processed for both immunofluoresence microscopy (a) and immunoblotting (b) 1 h later. The signals obtained by immunoblotting were quantified and the mean from three independent experiments calculated and plotted (c) against IR dose. GAPDH was used as a loading control. Both gammaH2AX and RAD51 increased linearly with IR dose. Signals for other markers of DNA repair, 53BP1, MRE11 and TP53, remained unchanged as assessed by immunoblotting (b). By immunofluoresence microscopy (a), MRE11 and 53BP1 redistributed into nuclear foci, particularly at the 2.28 Gy level. TP53 remained uniformly distributed throughout the nuclear compartment, but excluded from nucleoli (a). As the levels of gammaH2AX and RAD51 increased after exposure to IR (c), so the number of nuclear foci also increased (a). Scale bar, 10 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25924630), licensed under a CC-BY licence.

General chromatin effects of Adeltarod and Adeltaroddeltahbs. (c) No co-localization between Adeltarod structures and 53BP1 in HeLa cells indicates that chromatin is not damaged by the structures. Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/2073-4409/6/2/9), licensed under a CC-BY licence.

Specificity of antibodies for tryptophan 2,3-dioxygenase (TDO) by Western blot (A) and immunofluorescent staining (B). A: The total protein (20 μg) of the dentate gyrus, extracted from the wild-type (lanes 1 and 3) and Tdo-/- mice (lane 2), was subjected to 4% to 12% gradient sodium dodecyl sulfate polyacrylamide gel electrophoresis, and transferred to polyvinylidene difluoride membranes. TDO was detected as described in the Methods section. Note that a 45-kDa band was detected with the anti-TDO antibody. The positions of molecular weight markers are shown on the left. Lane 3: negative control without the primary antibody. beta -Tubulin is a positive control for the Western blot. B: The hippocampal sections of wild-type and Tdo-/- mice were stained with anti-TDO antibody. In sections of Tdo-/- mice, immunofluorescence signals were not detected in granule cells, interneurons (arrowhead), CA1, or CA3 cells, suggesting that the secondary antibody used in this study bound specifically to the primary antibody (mouse IgG). g, granule cell layer; h, hilus. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/20815922), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$CO attenuates mitochondrial dysfunction-derived sterile inflammation through TFEB-mediated mitophagy and mitochondrial biogenesis.Mice were exposed to LPS/D-galactosamine (D-GalN) with or without CO (250 ppm, 2 h/day) pretreatment, the blood and liver samples were collected 2 h and 6 h (b–i) after LPS/D-GalN administration. a The levels of ALT and AST were determined. Data are shown as mean ± SEM (n = 5). ***P < 0.001. b Liver sections were stained with H&E for morphological evaluation. c The levels of serum IL-1 beta were measured by ELISA. Data are shown as mean ± SEM (n = 4). **P < 0.01. ***P < 0.001. d Cytosolic mtDNA amounts in liver tissues were measured by qRT-PCR. Data are shown as mean ± SEM (n = 4). *P < 0.05. e Cleaved caspase-1 in cell lysates isolated from liver tissues was analyzed by immunoblotting. f The level of nuclear TFEB in liver tissues was determined by immunoblotting. Lamin B served as the nuclear standard. g Mitochondrial fractionation in liver tissues was analyzed by immunoblotting using antibodies against PINK1, Parkin, and LC3B-II. COX IV was used as marker for mitochondrial fractions. h, i The levels of autophagy-related genes and lysosomal genes (h) or mitochondrial biogenesis-associated genes (i) expressed in liver tissues were analyzed by qRT-PCR. Data are shown as mean ± SEM (n = 4). *P < 0.05, **P < 0.01. ***P < 0.001. j A scheme for the mechanisms of attenuating LSP/D-GalN-induced liver failure by CO through maintaining the balance between mitophagy and mitochondrial biogenesis. CO/CORM inhibits complexes of the mitochondrial respiratory chain (1) and then induces mitochondrial ROS (mtROS) production (2). Increased mtROS leads to PERK activation in the mitochondria-associated membrane (MAM) (3). The phosphorylation of PERK by mtROS increases intracellular calcium concentration. The activation of calcineurin by increased [Ca2 + ]i causes dephosphorylation of TREB (4), followed by its translocation into the nucleus. The nuclear translocation of TFEB increases the expression of genes involved in mitophagy (5) and mitochondrial biogenesis (6) Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30333475), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Precision quantitative image analysis workflow for the characterization of functional ITH.(a) Representative areas with positive immunohistochemical (IHC) staining for HIF-1 alpha , HIF-2 alpha , phospho-mTOR S2448, phospho-S6RP S235/236 and Ki-67. Only nuclear HIF-1 alpha and HIF-2 alpha expression was included in the analysis. Scale bar, 100 μm. (b) Overview of the quantitative IHC analysis process using a representative image of a whole-slide scanned pT1M1 tumour after phospho-mTOR S2448 staining. The image was overlaid with a virtual grid consisting of 1 mm2 squares that defines the tumour regions to be analysed. Non-cancerous tissue was subtracted manually (dark grey) and a positive pixel count was performed on the corrected area. Positivity per square was expressed as percentage of positive pixels. If a square contained non-tumorous tissue, the positivity was normalized to the corrected square area. Computed values were depicted as a heat map. Coloured squares represent tumour, whereas open squares indicate completely non-tumorous areas. Squares defined as peripheral tumour zone are marked by a black line. Scale bar, 1 cm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27291893), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reduction of HIF-1 alpha and HIF-2 alpha levels leads to differential effects on apoptosis in RCC10 cells. (A) RCC10 cells stably expressing wild-type VHLp30 or expressing a control shRNA targeting luciferase (Luc shRNA) or a pool of two shRNAs targeting either HIF-1 alpha or HIF-2 alpha were created. These cells were grown to confluence and lysed. Cell lysates were equally loaded and separated by SDS-PAGE. Western blots were performed for VHL, HIF-1 alpha , HIF-2 alpha , GLUT-1, and alpha -tubulin. (B) The RCC10 cell lines described in (A) were grown to confluence and treated with ultraviolet (UV) light and lysed one day later. Cell lysates were equally loaded and separated by SDS-PAGE. Western blots were performed for PARP, cleaved caspase-3, and alpha -tubulin. (C) The RCC10 cell lines described in (A) were glucose starved for 24 hours as described in Methods. Cell lysates were equally loaded and separated by SDS-PAGE. Western blots were performed for PARP, cleaved caspase-3, and alpha -tubulin. (D) The RCC10 cell lines described in (A) were serum starved for 2 days as described in Methods. Cell lysates were equally loaded and separated by SDS-PAGE. Western blots were performed for PARP, cleaved caspase-3, and alpha -tubulin. Similar results were seen with serum starvation for 4 and 6 days (data not shown). Image collected and cropped by CiteAb from the following open publication (//cancerci.biomedcentral.com/articles/10.1186/s12935-015-0175-3), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Expression of hypoxia-inducible factor (HIF) proteins after bilateral renal ischemia or sham ischemia. Immunoblots for HIF-1 alpha (A–C) and HIF-2 alpha (D–F) of tissue extracts from the cortex and outer and inner medulla of the left kidneys of rats 24 h and 5 days following recovery from either sham ischemia (○) or bilateral renal ischemia (●); n = 6 per group. G: typical image of the gel following electrophoresis. H: typical image of the nitrocellulose membrane following transfer. Values are expressed as medians (25th percentile, 75th percentile). Paired comparisons were performed using the Mann-Whitney U-test. Because paired comparisons were made at two time points, P values were conservatively adjusted using the Dunn-Sidak method with k = 2. PTr, PT, and PTr*T are the outcomes of two-way analysis of variance on ranking with the factors treatment (Tr) and time (T). AU, arbitrary unit; I1, 24 h after ischemia; I5, 5 days after ischemia; S1, 24 h after sham ischemia; S5, 5 days after sham ischemia. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30110566), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Active DNA end processing in late micronuclei compartments. a: Detection and quantification of RPA32 phosphorylation (pSerine 33) and ssDNA by specific antibodies in MN compartments of OVCAR-8 cells 36 h after 2-Gy irradiation. Note the nuclear signals have reduced while those in MN were intense. n = 3 biological repeats. Error bars = s.d. **P < 0.01; ***P < 0.005 (t-test). b: Immunostaining and enumeration of phosphorylated NBS1-pSerine 343, BRCA1-pSerine 1524 and TP53-pSerine 15 in MN compartments. **P < 0.01; ***P < 0.005 (t-test). c: Representative images (Upper panel) and quantification of RPA signal in MN compartments (Lower left) or RPA foci in nuclei (lower right) of post-IR OVCAR-8 cells after indicated treatments. Pro-resection genes (MRE11, BRCA1 and WDR70) were knockdown by specific siRNA. Percentage of positive RPA32-pS33 MN population was calculated after 36 h of IR treatment Image collected and cropped by CiteAb from the following open publication (//bmccancer.biomedcentral.com/articles/10.1186/s12885-018-4347-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Curcumin Analogs Induce Apoptosis in Cancerous Cells by Several Pathways. (a) E6-1 (Jurkat), dominant negative FADD (dnFADD) Jurkat, and overexpressing BCL-2 Jurkat were treated for 48 hours then stained for Annexin V and PI (b) E6-1 cells were plated and treated with or without the broad spectrum caspase inhibitor ZVAD(oMe)-FMK for 48 hours. Cells were stained for Annexin V and PI. Results were obtained using image-based cytometry with the Y-axis representative of percent of cells positive for Annexin V (green), PI (red), Annexin V and PI (yellow), or negative for both Annexin V and PI (blue). Values are expressed as a mean ± SD from three independent experiments. (c) E6-1 cells were treated for 24 hours with or without the broad spectrum caspase inhibitor ZVAD(oMe)-FMK and the studied compounds, lysed and subjected to Western blot analysis. (d) NHF and NCM460 cells were treated for 48 hours and 72 hours respectively, lysed and subjected to Western blot analysis. Bands were visualized with a chemiluminescence reagent. Images are representative of three independent experiments. Statistical calculations were performed using Two-Way ANOVA multiple comparison. *p < 0.05 vs % viable of Control (DMSO); #p < 0.05 vs % viable cells for groups without Z-VAD(oMe)FMK. 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.

PEM induces apoptosis of NSCLC cells via YIPF2-TNFRSF10B axis.a, b A549 (a) and H1792 (b) cells were treated with PEM at 5.0 μM for the indicated times (0, 6, 12, 24, 36 and 48 h). Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B and ACTB. c A549 cells were treated with doxorubicin (DOX) at various concentrations (0–2.0 μM) for 18 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B and ACTB. d Overexpression of YIPF2 in H1792 and H1299 cells in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B and ACTB. e Knockdown of YIPF2 expression by YIPF2–1 siRNA in A549 and H1792 cells in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B and ACTB. f Overexpression of YIPF2 in H1299 cells (left) or Knockdown of YIPF2 expression by YIPF2–1 siRNA in H1792 cells (right) in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10A and ACTB. g Three A549 cell lines (Ctrl, YIPF2, YIPF2 + siTNFRSF10B) in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B, CASP8, CASP3 and ACTB. h Three H1299 cell lines (Ctrl, siYIPF2–1, siYIPF2–1 + TNFRSF10B (short isoform)) in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B, CASP8, CASP3 and ACTB. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32303681), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Inhibition of CIP2A by siRNA impeded cell viability and DNA synthesis in HPV‐16E6–expressing cells. A, Elevated expression of CIP2A protein in 16E6‐expressing RPE1 cells. B, Western blot analysis of CIP2A and p53 proteins after transfection with scrambled siRNA (siCon) or CIP2A siRNA (siCIP2A) for 48 h. A representative of 3 independent experiments is shown. C, Cell viability assay of RPE1‐16E6 cells with CIP2A knockdown. D, Representative flow cytometry of BrdU staining profiles is shown. E, The mean and SD of BrdU‐positive cells from 3 experiments are summarized. **, 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.

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.

$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.$

Dynamics of the hypoxia-triggered decrease in MET phosphorylation and its reversal upon reoxygenation(A) MCF10A cells were incubated under normoxia or hypoxia for 1 h. They were then treated, under the same oxygen pressure, with 10 ng/mL HGF/SF for 5, 10, 20, 30, 60, 90 and 120 minutes. A control (Ctrl) without any HGF/SF stimulation was also performed. The same amount of protein was analyzed by western blotting with antibodies directed against: phosphorylated residues in the MET kinase domain, the MET kinase domain, the hypoxia marker HIF1a, phosphorylated Akt, Akt, Erk2, phosphorylated Erk, and actine. The positions of prestained molecular weight markers are indicated. Arrows indicate the positions of precursor and mature full-length MET. (B) MCF10A cells were incubated under hypoxia for 5, 10, 15 or 30 minutes. Another set of cells were incubated under hypoxia for 1 h and then returned to normoxia for 5, 10, 15 or 30 min (re-oxygenation). A control under normoxic (N) conditions was also included. The same amount of protein was analyzed by western blotting as previously described with the addition of GAB1 and its phosphorylated form. (C) MCF10A cells were placed under normoxic or hypoxic conditions for 1 h or hypoxia for 1 h and then normoxia for 10 minutes (reoxygenation). Cells were then treated at the indicated time with 10 ng/mL of HGF/SF. Cell lysates were incubated for AlphaScreen specific phospho-Erk and phospho-Akt quantitation. Error bars represent standard deviations (n = 3; ± SD). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29930749), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Downregulation of La autoantigen, hVAP-A, PSMA7, and NS5B leads to reduction in viral replication: overnight grown Huh-7.5 cells were transfected with La autoantigen, hVAP-A, and PSMA7 siRNAs for 48 h. Cells were then infected with the virus and again transfected with same siRNAs. After incubation total RNA was isolated and subjected to RT-PCR. NS5B siRNA was transfected once only after cells were infected. (a), (c), (e), and (g) represent RT-PCR results for target genes and NS5B gene upon treatment with La autoantigen, hVAP-A, PSMA7, and NS5B siRNAs, respectively. Densitometric analysis of (a), (c), (e), and (g) images is expressed in percentage in (b), (d), (f), and (h), respectively. The data represent mean ± standard deviation. ∗P value < 0.05 versus control considered statistically significant. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27446609), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

FUS does not bind VAPB or PTPIP51 in immunoprecipitation assays from transfected HEK293 cellsCells were transfected as indicated with control vector (CTRL), HA‐FUS + CTRL, myc‐VAPB + CTRL, or myc‐VAPB + either HA‐FUS, HA‐FUSR521C or HA‐FUSR518K. VAPB was immunoprecipitated via the myc‐tag and the samples probed on immunoblots for VAPB using rabbit VAPB antibody and for co‐immunoprecipitating FUS via the HA tag. Input VAPB and FUS were detected using myc and HA antibodies.Cells were transfected as indicated with control vector (CTRL), HA‐FUS + CTRL, HA‐PTPIP51 + CTRL or HA‐PTPIP51 + either HA‐FUS, HA‐FUSR521C or HA‐FUSR518K. PTPIP51 was immunoprecipitated using rat anti‐PTPIP51 and the samples probed for PTPIP51 using rabbit anti‐HA antibody and for co‐immunoprecipitating FUS using rabbit FUS antibody. Input PTPIP51 and FUS were detected using PTPIP51 and EGFP antibodies, respectively. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27418313), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

APCDD1 is co-regulated by MEIS1 and EWS-FLI1 and mediates the oncogenic role of MEIS1. (A, B) Silencing of (A) MEIS1 or (B) EWS-FLI1 downregulated expression of both APCDD1 mRNA and protein. Error bars represent mean ± SD of three replicates (**P < 0.01, ***P < 0.001). (C, D) APCDD1 knockdown potently (C) inhibited cell proliferation and (D) decreased colony formation. Error bars represent mean ± SD of three replicates (*P < 0.05, **P < 0.01, ***P < 0.001). (E) A673 and SKNMC cells stably expressing shMEIS1 were transiently transfected with plasmid encoding APCDD1, and subjected to immunoblotting and MTT assays. Bars represent mean ± SD of three replicates (**P < 0.01). (F) Proposed model showing that MEIS1 and EWS-FLI1 co-operatively activate APCDD1 transcription, thereby promoting the malignant phenotype of Ewing sarcoma cells. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30496486), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Pharmacologic inhibition of ATM rescues oxidative stress-induced senescence by suppressing ATM- and NEMO-mediated NF-kappa B activation. (A) Representative images of primary WT and Ercc1-/- MEFs were induced to undergo senescence by serial passaging at 20% oxygen. At passage 5, MEFs were grown in the presence or absence of KU-55933 (10 μM) for 72 hrs. Senescence was determined by SA-beta gal staining. Images were obtained at the magnification of 10x. (B) Quantitation of the percent SA-beta gal positive cells. Graph represents the mean +/- s.e.m. of three independent experiments. Student’s t-test, ***p <0.001, ****p <0.0001. (C) Passage 5 Ercc1-/- MEFs treated with vehicle or KU-55933 (10 μM) for 72 hours were collected and levels of p21 and p16INK4a were determined by western blotting. (D) Passage 5 Ercc1-/- MEFs were treated with KU-55933 (10 μM) for 72 hours and whole cell lysate (CL) and nuclear extracts (NE) were analyzed by immunoblotting for expression of proteins involved in the DNA damage response. (E) Whole cell lysate (CL) and nuclear extract (NE) were extracted from Ercc1-/- MEFs treated with 10 μM of KU-55933 for analysis of nuclear NEMO and p65. GAPDH was used as a loading control of total proteins and LaminA/C as a loading control of nuclear protein. (F) Passage 5 WT and Ercc1-/- MEFs transfected with a NF-kappa B-luciferase reporter construct were cultured in the presence or absence of KU-55933 (10 μM) and were collected for luciferase assays after 72 hours. (G) qRT-PCR analysis of mRNA expression in passage 5 WT and Ercc1-/- MEFs treated with or without of KU-55933 (10 μM) for 72 hrs. P values were determined using a Student’s t-test. *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/32201398), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Inhibition of splicing downregulates repair factors at both the mRNA and protein levels. (a) U2OS cells were treated with pladienolide B for 2, 6 and 16 h and irradiated (6 Gy, 1 h recovery) 1 h prior to termination of the treatment. The levels of the indicated mRNAs were measured through qPCR analysis. The change is relative to the DMSO control and two reference genes (18S rRNA and beta -actin). Means±S.D. are shown, n=3. (b) qPCR analysis of the levels mRNA and pre-mRNA for RNF8 and RAD51 in U2OS cells treated with pladienolide B or isoginkgetin for 2, 6 or 16 h and irradiated (6 Gy, 1 h recovery) 1 h prior to termination of this treatment. The change is expressed relative to the DMSO control value and the levels of mRNA for two reference genes (18S rRNA and beta -actin). Means±S.D. are shown, n=3. The arrows indicate the positioning of the PCR primers used and their sequences are shown in Supplementary Table S1. (c) Western blotting following pladienolide B treatment of U2OS cells as described in (a). beta -Actin was used as a loading control. Densitometric quantification of each protein is shown in Supplementary Figure S2B. The three MDC1 bands correspond to the unphosphorylated, phosphorylated and hyperphosphorylated forms of full-length MDC1 Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27315300), 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.

Lysyl Oxidase (LOX) expression in malignant and non-malignant human prostate tissue.(A) Representative immunohistochemical staining of LOX (brown) in sections of non-malignant prostate tissue specimens (TINT epithelium and TINT stroma) from two prostate cancer patients (original magnifications x 200). Case A show weak epithelial staining (score 1) and Case B strong epithelial staining (score 3). (B) Consecutive sections from non-malignant prostate tissue stained with in situ hybridization or immunohistochemistry for LOX. Note that mRNA (brown dots) and protein (brown) expression in individual glands were related, in the stroma in contrast mRNA was generally low but protein often detected (original magnifications x200). (C) LOX mRNA expression in non-malignant prostate tissue (TINT) (n = 12), in primary prostate tumor tissue (n = 12), and in castration-resistant bone metastases (CRPC) (n = 30) using Illumina gene expression array. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/26501565), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Recruitment of 53BP1 and TRF2 to fs NIR laser-induced DNA lesions. (A) EGFP-53BP11220-1711 and TRF2-YFP cell lines were subjected to NIR laser microirradiation with the input power ranging from 20 mW to 155 mW (corresponding to ≈7 × 1010 W/cm2 to 5.41 × 1011 W/cm2 peak irradiance) as indicated. EGFP-53BP11220-1711 and TRF2-YFP recruitment was monitored for 15 min and 6 min p.i., respectively. The number of the cells with the recruitment and the number of cells examined are shown at the top of each histogram. Detailed microirradiation energy and irradiance calculations versus the laser input power (the power of the laser beam entering the microscope) are shown in Table 2. (B) Time course fluorescence measurement of the damage-site recruitment of EGFP-53BP11220-1711 and TRF2-YFP. PtK2 cells stably expressing EGFP-53BP11220-1711 or TRF2-YFP were damaged with either low (60 mW input power with peak irradiance of ≈2.1 × 1011 W/cm2; upper panel) or high (100 mW input power with peak irradiance of 3.49 × 1011 W/cm2; lower panel) dose of NIR laser microirradiation as indicated. For EGFP-53BP11220-1711, four and three cells were followed for 30 min for 60 mW and 100 mW damage, respectively. For TRF2-YFP, four cells each were followed for 4 min. The fluorescent intensity at the damage site was divided by that in the nucleoplasm and was subtracted by one for normalization. (C) Similar experiments as in (B) were performed following EGFP-53BP11220-1711 recruitment, and live fluorescent images of one representative cell each at indicated p.i. time points are shown. Time 0:00 refers to pre-laser condition. The irradiation site is indicated by a black line at time 0:00 and white arrowheads. Consistent results were obtained with 12 and 8 cells examined for the low (60 mW) and high (100 mW) irradiation condition, respectively. Scale bar = 10 μm. (D) Similar experiments as in (C) for TRF2-YFP. Consistent results were obtained with 11 and 12 cells examined for the low (60 mW) and high (100 mW) irradiation condition, respectively. Scale bar = 10 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/26424850), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Bt, Bv, Ca, Fi, Gt, Pm, Po, Pm, Fe, Pm-Or, Rb, Sh

Species Glossary

Reactivity: Hu, Mu, Rt, Bt, Bv, Ca, Fi, Gt, Pm, Po, Pm, Fe, Pm-Or, Rb, 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	NB100-304AF350
Alexa Fluor 405	NB100-304AF405
Alexa Fluor 488	NB100-304AF488
Alexa Fluor 532	NB100-304AF532
Alexa Fluor 594	NB100-304AF594
Alexa Fluor 647	NB100-304AF647
Alexa Fluor 700	NB100-304AF700
Alexa Fluor 750	NB100-304AF750
Allophycocyanin	NB100-304APC
Allophycocyanin/Cy7	NB100-304APCCY7
Biotin	NB100-304B
DyLight 350	NB100-304UV
DyLight 405	NB100-304V
DyLight 488	NB100-304G
DyLight 550	NB100-304R
DyLight 594	NB100-304DL594
DyLight 650	NB100-304C
DyLight 680	NB100-304FR
DyLight 755	NB100-304IR
FITC	NB100-304F
HRP	NB100-304H
Janelia Fluor 549	NB100-304JF549
Janelia Fluor 646	NB100-304JF646
PE	NB100-304PE
PE/Atto594	NB100-304PEATT594
PE/Cy5.5	NB100-304PECY55
PE/Cy7	NB100-304PECY7
PerCP	NB100-304PCP
mFluor Violet 450 SE	NB100-304MFV450
mFluor Violet 500 SE	NB100-304MFV500
mFluor Violet 610 SE	NB100-304MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-304SS

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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.

Reactivity:

Hu, Mu, Rt, Ca, Fe, Po

Species Glossary

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

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

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Sample Size Available

Sizes: Sample Size Available

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Unconjugated

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NB600-408

Collagen I Antibody

(16 Reviews)

(298 Publications)

0.1 mg

NB600-408

Rat colon tissue stained with Collagen I antibody (red) and Hoechst (blue). IHC-P image submitted by a verified customer review.

<b>Biological Strategies Validation. </b>Myofibroblast differentiation requires CD271/NGF activation. Representative alphaSMA (red) and COLLAGEN 1 (red) immunostaining with DAPI (blue) in non-treated (control), TGFbeta-treated cells with or without gamma-secretase inhibitor. Scale bars: 50 um Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-019-09992-3) licensed under a CC-BY license.

Lane 1: Human Collagen Type 1. Lane 2: None. Load: 50 ng per lane. Primary antibody: Collagen Type I antibody at 1:1000 overnight at 4C. Secondary antibody: DyLight 649 rabbit secondary antibody at 1:20000 for 30 min at RT. Block: incubated with blocking buffer for 30 min at RT. Predicted/Observed size: 139 & 130 kDa, 139 & 130 kDa for Collagen Type I. Other Band(s): Collagen Type I splice variants and isoforms.

Representative Western blot image for Collagen I, Collagen 2 and GAPDH. N/group = 8. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1111/jcmm.12367) licensed under a CC-BY license.

Macro- and micro-architecture of human adipose tissue lobule. Representative immunostaining of the human AT lobule with Collagen I (COL1), CD34, and DAPI. The position of the septa is underlined, scale bar: 100 um. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-019-09992-3) licensed under a CC-BY license.

Primary human cardiac fibroblast cells were stained with anti-Collagen I antibody. Cells were cultured for 3 days in DMEM with 10% fetal calf serum. ICC/IF image submitted by a verified customer review.

Cell lysate from human trabecular meshwork. Antibody at 1:1.000. WB image submitted by a verified customer review.

Patient-derived CAF and NF CDMs are architecturally and functionally distinct. (e) Collagen I (red) and fibronectin (blue) staining of patient #1 and #3 NF and CAF CDM. Scale bar, 20 μm. Image collected and cropped by CiteAb from the following publication (//www.nature.com/doifinder/10.1038/ncomms12237) licensed under a CC-BY license.

Staining of human HaCaT-ras A5-RT3 (SCC) frozen tumor sections. Image provided by Dr. Wa'el Al Rawashdeh of RWTH Aachen University.

Livers were harvested at 9 wpi, formalin-fixed, and then cryosectioned and stained for markers of inflammation and liver fibrosis. Maximum intensity projections of 12-17 um thick liver sections with immunofluorescently labeled nuclei (DAPI white) alpha-smooth muscle actin NBP2-34760APC (yellow), CD45 (red) and Collagen I NB600-408 (top) or Collagen III/COL3A1 NB600-594 (bottom) (blue) in the liver of uninfected or 9 wpi WT mice. Scale bars represent 50 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32973293/) licensed under a CC-BY license.

Analysis in porcine burn wound lysate using anti-Collagen I alpha 1 antibody. WB image submitted by a verified customer review.

Detection of collagen I in Wistar rat hepatic stellate cells (HSC) in control (GFP-transduced) (left lane) and PPARg-transduced cell lysates (right lane). Protein staining shown below each blot depicts equal protein loading. An equal amount of the whole cell protein (100 ug) was separated by SDS-PAGE and electroblotted to nitro-cellulose membranes. Proteins were detected by incubating the membrane with anti-Collagen I antibody at a concentration of 0.2-2 ug/10 mL in TBS (100 mM Tris-HCl, 0.15 M NaCl, pH 7.4) with 5% non-fat milk. Detection occurred by incubation with a HRP conjugated secondary antibody at 1 ug/10 mL. Proteins were detected by a chemiluminescent method using the PIERCE ECL kit (Amersham Biosciences).

Balb/3T3 mouse embryonic fibroblast cell line stained with Collagen I antibody. IHC-P image submitted by a verified customer review.

Mouse pancreas tissue stained with Collagen I antibody. IHC-P image submitted by a verified customer review.

Human lung tissue section stained with Collagen I antibody at 1:400.

FFPE right lobe of the liver tissue section. A:Central Vein (CV) fibrosis, B: Non-fibrotic CV, C: Perisinusodial fibrosis, D: Non-fibrotic area, E: Protat tract fibrosis, F: Septal fibrosis (arrow). Antigen retrieval: not required. Primary antibody: Anti-collagen type I at 1:1250 for 4C for 24hr. Secondary antibody: Peroxidase biotin-streptavidin rabbit secondary antibody at 1:10,000 for 45 min at RT. Localization: Anti-collagen type I is intra and extracellular. Staining: 3.3'-diaminobenzidine tetrahydrochloride was used as the chromogen. Nuclei were counterstained purple with hematoxylin.

Analysis of FFPE human lung. Primary antibody: Collagen I 1:400. Secondary antibody: Peroxidase goat anti-rabbit at 1:10,000 for 45 min at RT. Localization: Strong staining was observed in the extracellular matrix of the lung. Epithelial cells were negative. Staining: antibody as precipitated red signal with a hematoxylin purple nuclear counterstain.

Analysis of HRP conjugate of NB600-408. FFPE Human Skin at pH 6. Primary antibody: Collagen Type I antibody at 10 ug/mL for 1 h at RT. Localization: Collagen Type I is secreted in the extracellular matrix.

Analysis of HRP conjugate of NB600-408. FFPE Human Skin tissue at pH9. Primary antibody: Collagen Type I antibody at 10 ug/mL for 1 h at RT. Localization: Collagen Type I is secreted in the extracellular matrix. Staining: Collagen Type I as precipitated brown signal (A) with hematoxylin purple nuclear counterstain. With corresponding negative conrol (B).

Imaging of rat calvarial defect bone. IHC-P image submitted by a verified customer review.

Immunohistological staining of EG treated rat kidneys for Collagen. Magnification X40, Scale bar 100 um. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0185009), licensed under a CC-BY license.

Analysis of Collagen I Antibody on alcohol-fixed rat lung tissue. Primary antibody dilution 1:200. Image from verified customer review.

Immunohistochemistry of Collagen I antibody. Tissue: human lung. Fixation: formalin fixed paraffin embedded. Antigen retrieval: user optimized. Primary antibody: Collagen 1 1:400 Secondary antibody: Peroxidase goat anti-rabbit at 1:10,000 for 45 min at RT. Localization: Strong staining was observed in the extracellular matrix of the lung. Epithelial cells were negative. Staining: antibody as precipitated red signal with a hematoxylin purple nuclear counterstain.

Immunohistochemistry of Rabbit Anti-collagen type I antibody. Tissue: right lobe of the liver section. A:Central Vein (CV) fibrosis, B: Non-fibrotic CV, C: Perisinusodial fibrosis, D: Non-fibrotic area, E: Protat tract fibrosis, F: Septal fibrosis (arrow). Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: Anti-collagen type I at 1:1250 for 4C for 24hr. Secondary antibody: Peroxidase biotin-streptavidin rabbit secondary antibody at 1:10,000 for 45 min at RT. Localization: Anti-collagen type I is intra and extracellular. Staining: 3.3'-diaminobenzidine tetrahydrochloride was used as the chromogen. Nuclei were counterstained purple with hematoxylin.

Immunohistochemistry-Paraffin: Rabbit Polyclonal Collagen I Antibody [NB600-408] - Collagen I was stained in human FFPE tissue. Primary antibody dilution: 1/1000 in blocking buffer. HIER antigen retrieval at pH 9 for 20min. AF750 conjugated version of the antibody was used (Catalog # NB600-408AF750). Image from a verified customer review.

Effects of fasudil treatment on tumor stroma in KPC mice.A) Pancreatic tumor tissues from vehicle, gemcitabine, or combination of gemcitabine plus fasudil treated KPC mice were harvested and stained for various stromal markers. Representative images are shown of H&E staining, alpha -SMA, Desmin, CD31, Collagen I, and Movat's pentachrome staining.

Western blot analysis. Comparison of collagen 1 (A and B) and collagen 2 (D and E) among the anterior horn, the body and the posterior horn: (A and D) young model; (B and E) adult model. Comparison of collagen 1 (C) and collagen 2 (F) content between the young and the adult specimens. Values with different superscripts differ for P 50 cells. G, histograms show the rate of colony formation. Data represent the average of three experiments (mean ± SD). **P < 0.01, compared to DMSO treated cells. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/25204354), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Phd2 deletion leads to activation of the Akt–mTOR pathway. a RPPA analysis of tumors with homozygous deletion of Phd2. Tumor tissues from Tyr::CreER; BRafV600E; Phd2−/− or Tyr::CreER; BRafV600E mice were processed and analyzed by RPPA assays. The analyses identified proteins that were significantly changed in mouse melanomas compared to nevi. b Activation of Akt–mTOR pathway after phd2 deletion. Tumor tissues were processed and western blots showed stabilization of HIF-1 alpha and HIF-2 alpha proteins after Phd2 depletion. Increased phosphorylation of Akt, 4EBP1 and S6K was observed in tumors from Tyr::CreER; BRafV600E; Phd2−/− compared with those of Tyr::CreER; BRafV600E mice. c Re-expression of Phd2 inhibits the Akt–mTOR pathway. A BRafV600E; Phd2−/− mouse melanoma cell line was established from melanomas in Tyr::CreER; BRafCA; Phd2lox/lox mice. Phd2 was ectopically reintroduced in these tumor cells. Western blot analysis showed that degradation of HIF-1 alpha and HIF-2 alpha proteins with decreased expression of VEGFR2 decreased phosphorylation of Akt, 4EBP1 and S6K. d Pharmacological inhibition (FM19G11) of HIF pathway in BRafV600E; Phd2−/− melanoma cells. A similar but more pronounced inhibition of the Akt–mTOR pathway was observed using the HIF inhibitor. beta -Actin was used as a loading control. Results are representative of three independent experiments Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30575721), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Aberrant HIF1 alpha expression in PDAC cancer associated fibroblastsA. The level of MCT4 was determined in the indicated established pancreatic cancer cell lines and CAF cultures by immunoblotting. B. The level of MCT1 was determined in the indicated established pancreatic cancer cell lines and CAF cultures by immunoblotting. C. HIF alpha and MCT4 levels were determined by immunoblotting following the knockdown of HIF1 alpha with RNAi or the induction of hypoxia through the use of CoCl2. D. The stability of HIF1 alpha was evaluated in CAFs following treatment with cycloheximide. E. HIF1 alpha was detected by immunoblotting in the presence of cycloheximide in the absence or presence of the proteasome inhibitor MG132. F. HIF1 alpha protein levels were determined in the absence and presence of DMOG. G. RNA levels of the indicated genes as determined by microarray analyses (**p < 0.01 for tumor cell line vs. CAF). H. MG132 was used to interrogate the synthetic rate of HIF1 alpha protein in the indicated cell lines. I. The expression of HIF1 alpha target genes was evaluated in the indicated cell lines by microarray analysis (**p < 0.01 for tumor cell vs. CAF). 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.

Chromatin opening activates DDR via a mechanism that requires catalytically functional LOXL2 but can be independent of DNA damage. a gamma -H2AX and 53BP1 staining and foci quantification are shown by immunofluorescence with a specific antibody for gamma -H2AX (left image) or for 53BP1 (right image). Dot graphs indicate the number of foci for gamma -H2AX (upper graph) and 53BP1 (lower graph) per cell in control and LOXL2 KD conditions. b gamma -H2AX staining and foci quantification are shown by immunofluorescence with the indicated antibody after LOXL2 reinfection. MDA-MB-231 cells were first infected with control or LOXL2 KD lentivirus and then, after puromycin selection, again with GFP (MOCKGFP+), wild-type LOXL2-IRES-GFP (LOXL2GFP+), or LOXL2mut-IRES-GFP (LOXL2mutGFP+). Cells were fixed after 24 h. Dot graphs indicate the number of gamma -H2AX foci per GFP-positive cells containing MOCKGFP+ (upper graph), LOXL2GFP+ (middle graph), or LOXL2mutGFP+ (lower graph). c Dot graphs indicate the number of gamma -H2AX (upper graph) and 53BP1 (lower graph) foci per cell in control and LOXL2 KD cells after treatment with 200 μM cordycepin for the indicated timepoints. d gamma -H2AX and 53BP1 staining and foci quantification are shown after immunofluorescence with the indicated antibodies in non-replicative conditions. Dot graphs indicate the number of the gamma -H2AX (left graph) and 53BP1 (right graph) foci in control and LOXL2 KD cells. e Representative image showing DNA damage in control or LOXL2 KD MDA-MB-231 cells, visualized by the alkaline comet assay. Cells were treated with 0.3 μM doxorubicin for 24 h. The graph shows the percentage of MDA-MB-231 positive cells. f Chromosome alterations in control and LOXL2 KD MDA-MB-231 cells. Error bars indicate the SD from at least three independent experiments. *P < 0.05, **P < 0.01, and ***P < 0.001, ****P < 0.0001 Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31462706), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Insuline-like growth factor (IGF1) antagonized the effects of miR-106a-5p on myogenic differentiation in C2C12. All data were collected from C2C12 myotubes 5 d post differentiation. (A) MyHC immuno-staining of cells incubated in differentiation medium with or without 75 μM IGF1 recombinant protein. Scale bars = 200 μm; (B–D) The statistical results of differentiation index, fusion index and the populations of myotubes, respectively; (E) The expression of miR-106a-5p upon IGF1 stimuli; (F) The mRNA levels of MyoD, MyoG, MyHC in myoblasts cultured in differentiation medium containing 75 μM IGF1 recombinant protein or not; (G) Western-blot analysis of myogenic regulatory factors (MyoD, MyoG, MyHC) and key molecules in PI3K-AKT pathway in cells; (H,I) The statistical results of Figure 3G; (J) Immunostaining of MyHC (red) and DAPI (blue). Scale bars = 200 μm; (K–M) The statistical results of differentiation index, fusion index and the populations of myotubes, respectively; (N) The mRNA levels of MyoD, MyoG, MyHC; (O) Western blot analysis for MyoD, MyoG, MyHC and p-AKT/AKT upon IGF1 challenge; (P) Statistical analysis of MyoD, MyoG, MyHC and p-AKT/AKT levels in Figure 3O. Data were presented as mean ± SEM. n = 3 per group. * p < 0.05, ** p < 0.01. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30004470), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

LukAB is a potent activator of Caspase 1.(A) THP1 cells were intoxicated with 50 ng/mL LukAB for 1 hour and cell lysates were analyzed by immunoblot for Caspase 1 cleavage, which indicates activation. (B and C) THP1 cells were incubated with FLICA-1 (660-YVAD-FMK) then intoxicated with culture filtrates (1% v/v) from the indicated S. aureus stains. After 1 hour, cells were washed, fixed and analyzed by flow cytometry. Panel B shows a representative flow plot from one experiment, and panel C shows the corresponding mean florescence intensities (MFI). (D) THP1 cells were incubated with FLICA-1, intoxicated with purified LukAB (50 ng/mL) for 1 hour, then washed, fixed and analyzed by flow cytometry. (E) FLICA-1 was added to THP1 cells then LukAB (50 ng/mL) was added for time-course samples in descending order. All samples were washed and fixed at the same time, corresponding to different LukAB incubation times, then analyzed by flow cytometry. THP1 cells (F) and primary CD14+ human monocytes (G) were incubated with FLICA-1 and intoxicated with a dose titration of the indicated purified S. aureus leukotoxins for 1 hour. Cells were washed, fixed and analyzed by flow cytometry. Bars represent the standard error of the mean of triplicate samples. EC50 values are also shown. Each graph is representative of three experiments. Primary cell experiments include three independent donors. Asterisks indicate significance at a p-value of ≤ 0.05 by Tukey’s multiple comparisons post-test for 1-way or 2-way ANOVA, as appropriate. Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.ppat.1004970), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Glucose fluctuations promoted the activation of NLRP3 inflammasome. (A) Immunofluorescence staining of NLRP3-ASC interaction in rat hearts. The heart sections were labeled with anti-NLRP3 (red), anti-ASC (green), and DAPI (blue) (n = 3 per group). (B–D) The mRNA levels of NLRP3, IL-1 beta and IL-18 in rat hearts of the three groups (n = 4 per group). (E–H) The protein expressions of NLRP3, ASC and cleaved caspase-1 in rat hearts of the three groups (n = 4 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.

EPS15 is targeted via a SPOP/SPOPL binding consensus motif.(A) Cartoon of human EPS15 domain-organization and the amino-acid sequence. Indicated by color code are the SPOP/SPOPL binding site (red) and the lysine residue (yellow), which is ubiquitinated in a CRL3SPOPL–dependent manner in vivo. In addition, the amino-terminal Ca2+-binding EF-hand motifs (EH), the coiled-coil domain involved in dimerization and the two carboxy-terminal ubiquitin-interacting motifs (UIMs) involved in ubiquitin-binding are indicated. (B) EPS15 ubiquitin-profiling. Peptides containing EPS15 modification sites were quantified with LC-MS/MS after enrichment of the K-epsilon -GG motif from whole cell digests of HeLa cells treated with siSPOPL or siControl. Normalized precursor mass intensity profiles for EPS15 sites corresponding to K793, K801 and K693 are shown (raw data in Figure 4—figure supplement 1B). Quantification of the beta -Actin K113 and the polyubiquitin K11 linkage peptide control for comparable enrichment. Data are mean ± SD, N = 3. **p≤0.01. (C) Purified SPOPL was incubated as indicated with GST-tagged wild-type EPS15 or GST-EPS15 mutants, where the predicted SPOPL binding motifs have been mutated individually (GST-EPS15S605-S607A and EPS15S744-S746A, respectively), pulled down with glutathione sepharose (IP [GST]) and bound proteins were analyzed by Coomassie blue staining (upper panel) and immunoblotting (lower panels). Note that SPOPL readily binds to GST-EPS15 and GST-EPS15S605-S607A, but this interaction is strongly reduced with the GST-EPS15S744-S746A mutant. (D) HeLa cells stably expressing GFP-tagged wild-type EPS15, the EPS15S744-S746A or the EPS15K793R mutants from a doxycycline-inducible promoter were transfected as indicated (+) with control siRNA or siRNA depleting SPOPL. The levels of EPS15-GFP, EGFR and for control tubulin (TUB) were analyzed by immunoblotting with specific antibodies. Experiments were quantified in Fiji and the EPS15 levels plotted as fold-increase compared to controls. Data are mean ± SEM, N = 4. *p≤0.05. Note that SPOPL depletion does not further increase the levels of both EPS15 mutants. (E) Total cell extracts were prepared from HeLa cells expressing either GFP-tagged wild-type, the EPS15S744-S746A mutant or the EPS15K793R mutant in the presence (+) or absence (-) of HA-tagged SPOPL overexpression. The levels of EPS15-GFP, SPOPL-HA and control GAPDH were analyzed by immunoblotting. Note that overexpression of SPOPL-HA is able to induce degradation of wild-type but not the EPS15S744-S746A-GFP or the EPS15K793R-GFP mutant.DOI://dx.doi.org/10.7554/eLife.13841.009EPS15 is targeted via a SPOP/SPOPL binding consensus motif.(A) Alignments of the carboxy-terminal domains of EPS15 proteins from various species. Conserved SPOPL-binding motifs and putative ubiquitination sites are highlighted by yellow boxes. (B) Peptides containing EPS15 modification sites were quantified with LC-MS/MS after enrichment of the K-epsilon -GG motif from whole cell HeLa digests treated with siSPOPL and siControl. Raw intensities for each of the triplicate LC-MS/MS runs are shown with each of the siControl conditions scaled to 100% intensity. Normalized precursor mass intensity profiles for EPS15 sites corresponding to K793, K801 and K693 are shown, with only K793 showing significant downregulation in the depletion condition. Quantification of a peptide corresponding to beta -Actin K113 and the poly-ubiquitin K11 linkage peptide is also shown to demonstrate that enrichment variations did not influence the quantification of the EPS15 sites. Additionally, the total ion chromatographic intensities for each run are plotted to provide insight into the consistency of each of the separate experiments performed on different days. Data are mean ± SD, N = 3. (C) HeLa cell lines stably expressing wild-type EPS15-GFP, the EPS15S744-746A-GFP mutant or the EPS15K793R-GFP mutant from the inducible doxycycline-promoter were treated with doxycycline for 3 days, and analyzed by live cell imaging. Displayed are maximal projections of Z-stack acquisitions, fully covering cell height. Scale bar = 10 μm.DOI://dx.doi.org/10.7554/eLife.13841.010 Image collected and cropped by CiteAb from the following open publication (//elifesciences.org/articles/13841), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Examples of PNUTS staining: (a) comparison of PNUTS staining in (1) normal and (2) cancerous prostate glands of the same TMA spot. Cancer spots with (b) lack of staining and (c) weak, (d) moderate, and (e) strong staining. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32377272), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

The mRNA and protein expression of cytochrome c oxidase 4 (COX4), toll-like receptor (TLR) 9, myeloid differentiation factor 88 (MyD88), and nuclear factor (NF)-kappa B in lung tissues from spontaneous breathing group (CON group), normal tidal volume (NTV) group, high tidal volume (HTV) group, chloroquine (CQ) pretreatment upon HTV stimulation group (HTV + CQ). (A) Levels of COX4, TLR9, MyD88, and NF-kappa B mRNA. (B) Levels of COX4, TLR9, MyD88, and NF-kappa B protein by Western blot. (C) Relative expression of COX4, TLR9, MyD88, and NF-kappa B protein. Fold expression for target genes was normalized to that measured for the beta -actin gene. Both of these experiments were in triplicate. aP < 0.05 vs. CON group; bP < 0.05 vs. NTV group; and CP < 0.05 vs. HTV group. Image collected and cropped by CiteAb from the following open publication (//www.frontiersin.org/article/10.3389/fimmu.2018.01477/full), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Interaction of Nesprin-2-SMC and Nesprin-2 with SMC2 and SMC4. (a) Precipitation of SMC2 and SMC4 with GST-Nesprin-2-SMC from HaCaT cell lysates. Precipitates were resolved on SDS-polyacrylamide gels (10% acrylamide) and probed with SMC2 and SMC4 specific antibodies. SPN, supernatant after pulldown; PD, pulldown. The lower molecular weight band in the SMC2 pulldown is presumably a breakdown product. (b) Immunoprecipitation of SMC2 from HaCaT cell lysates with Nesprin-2 specific mAbK20-478 and of Nesprin-2 with SMC2 specific antibodies. GFP-specific monoclonal antibodies were used for control. The antibodies used for immunoprecipitation are indicated above the panels (IP). The blots were probed with the antibodies listed on the right (WB). Immunoprecipitates were resolved on gradient gels (3–12% acrylamide) and 10% acrylamide gels as appropriate. The data are from one blot; however, the input was not directly adjacent to the SMC2 IP. (c) Interaction of CAP-H2 (condensin II) and CAP-H (condensin I) with Nesprin-2-SMC. Pulldowns were performed with HaCaT cell lysates and GST for control and GST-Nesprin-2-SMC as indicated. Unsynchronized cells were used for the experiments shown in (a)–(c). (d) Analysis of the Nesprin-2-SMC interaction with SMC2 during the cell cycle. HaCaT cells were synchronized with RO-3306 or other reagents as described in Materials and Methods in order to obtain the relevant cell cycle phases. Cell cycle phases were assessed by FACS analysis; the results are depicted in the accompanying diagram. Pulldown was carried out with GST-Nesprin-2-SMC bound to GST-Sepharose. GST was used for control. The blot was probed with SMC2 specific antibodies. (e) Localization of Nesprin-2 as detected with mAb K81-116-6 (green) during mitosis in HaCaT cells. DNA was stained with DAPI. Arrow points to filamentous staining across the chromosomes. (f) Nesprin-2 distribution in HaCaT cells during mitosis as detected with mAb K20-478 (green) and pAbK1 (red). DNA was detected with DAPI. Bar, 10 μm. (g) Nesprin-2 presence on chromosomes. Different Z-stacks (from top to bottom: 0 μm, 0.21 μm 0.42 μm, and 0.84 μm) from a COS7 cell in anaphase stained with mAb K20-478. DNA was stained with DAPI. Bar, 5 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29445399), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Quantification of knockdown of beta 1 integrin in hS/PC microstructures. hS/PC microstructures formed in HA-based hydrogels and were transfected with scrambled siRNA or ITGB1 siRNA. Immunocytochemistry was used to determine beta 1 integrin expression and spatial distribution of beta 1 integrin 46 h post-transfection. Image analysis software was used to quantify beta 1 integrin expression at microstructure surfaces from confocal micrographs and coverage indices were calculated for scrambled siRNA (A) and ITGB1 siRNA (B) groups where a 63% reduction in beta 1 integrin expression at microstructure surfaces was quantified (C). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31750298), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Bv, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, 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	NB600-408AF350
Alexa Fluor 405	NB600-408AF405
Alexa Fluor 488	NB600-408AF488
Alexa Fluor 532	NB600-408AF532
Alexa Fluor 594	NB600-408AF594
Alexa Fluor 647	NB600-408AF647
Alexa Fluor 700	NB600-408AF700
Alexa Fluor 750	NB600-408AF750
Allophycocyanin	NB600-408APC
Biotin	NB600-408B
DyLight 350	NB600-408UV
DyLight 405	NB600-408V
DyLight 488	NB600-408G
DyLight 550	NB600-408R
DyLight 594	NB600-408DL594
DyLight 650	NB600-408C
DyLight 680	NB600-408FR
DyLight 755	NB600-408IR
FITC	NB600-408F
HRP	NB600-408H
Janelia Fluor 549	NB600-408JF549
Janelia Fluor 646	NB600-408JF646
PE	NB600-408PE
PerCP	NB600-408PCP

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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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NB600-308

GFP Antibody

(18 Reviews)

(262 Publications)

0.1 mg

NB600-308

Analysis of GFP in transgenic mouse pancreas in OCT. The blue region in the center is an islet which does not express GFP and is negative. Image from verified customer review.

Analysis of GFP tagged PKA subunits over-expressed in HEK293T cells using GFP antibody. Image from verified customer review.

Staining of GFP+ hepatocytes (brown) transplanted into the liver of a recipient animal using anti-GFP antibody. Cyclin D1 double staining in blue. Image from verified customer review.

Total cell lysates (20ug) with protein of interest tagged with GFP was loaded. Blotted with GFP antibody [HRP] NB600-308H, without any secondary antibody. Image from verified customer review.

Germaria from wt or chico1/KG mutant ovaries endogenously expressing Myc-GFP stained with anti-GFP, anti-1B1, and DAPI. Scale bars, 10 um. (F) Quantification of relative Myc-GFP intensity in germarium region 2B from wt or chico1/KG mutant ovaries. n = 10 germaria for each genotype. Error bars represent SEM. *p 0.05), as determined by Student’s t-test. Representative blots for NANOG, OCT4, SOX2 and GAPDH are shown below the graphs d. MM – molecular weight marker, C – tissues obtained from control cows, ADENO – tissues obtained from cows with adenomyosis Image collected and cropped by CiteAb from the following open publication (//www.rbej.com/content/13/1/110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Protein expression and nuclear foci formation in breast cancer cell lines. (a) Cellular protein levels. 53BP1 and Vimentin band intensities were normalized to loading controls. Values for T47D cell extracts, loaded as a reference on each blot, were set to 1. Mean relative Western blot signals per cell line were calculated from two independent experiments. Box plots in the upper panels show mean values (cross), the median (line), and 95% CI (whiskers) for untreated and for 7d Eribulin-treated non-TNBC (N=4) and TNBC cell lines (N=7). The Kruskal–Wallis test did not reveal significant differences in the case of 53BP1. Representative immunoblots are shown in the bottom panels. Vimentin staining was detected in all TNBC but was negative in all non-TNBC cell lines in which BT20 served as a reference. Actin normalized protein levels are indicated above the 53BP1 and Vimentin signals. (b) Focal accumulation of 53BP1 in the nucleus of MBC cell lines. 53BP1 foci were detected by immunofluorescence microscopy in MBC cell lines. The graph presents quantitative data in box plots as in A; ~300–900 nuclei per sample were scored in two independent experiments each, and mean values were calculated for each cell line. *P<0.05, Kruskal–Wallis test, Mann–Whitney U test. Representative immunofluorescence images are displayed in Supplementary Figure S3. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32283863), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

All-NA

Species Glossary

Reactivity: All-NA

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB600-308AF350
Alexa Fluor 405	NB600-308AF405
Alexa Fluor 488	NB600-308AF488
Alexa Fluor 532	NB600-308AF532
Alexa Fluor 594	NB600-308AF594
Alexa Fluor 647	NB600-308AF647
Alexa Fluor 700	NB600-308AF700
Alexa Fluor 750	NB600-308AF750
Allophycocyanin	NB600-308APC
Allophycocyanin/Cy7	NB600-308APCCY7
Biotin	NB600-308B
DyLight 350	NB600-308UV
DyLight 405	NB600-308V
DyLight 488	NB600-308G
DyLight 550	NB600-308R
DyLight 594	NB600-308DL594
DyLight 650	NB600-308C
DyLight 680	NB600-308FR
DyLight 755	NB600-308IR
FITC	NB600-308F
HRP	NB600-308H
Janelia Fluor 549	NB600-308JF549
Janelia Fluor 646	NB600-308JF646
PE	NB600-308PE
PE/Atto594	NB600-308PEATT594
PE/Cy5.5	NB600-308PECY55
PE/Cy7	NB600-308PECY7
PerCP	NB600-308PCP
mFluor Violet 450 SE	NB600-308MFV450
mFluor Violet 500 SE	NB600-308MFV500
mFluor Violet 610 SE	NB600-308MFV610

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NB300-109

Tyrosine Hydroxylase Antibody

(7 Reviews)

(219 Publications)

0.1 ml

NB300-109

Tyrosine hydroxylase immunoreactivity in the central complex and the lateral accessory lobe. A-D: Frontal sections (immunoperoxidase preparations, dorsal to the top). E: Horizontal section (immunofluorescent preparation, posterior to the top). Scale bars = 100 um. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0160531), licensed under a CC-BY license.

Dopamine neurons in the mouse substantia nigra. ICC/IF image submitted by a verified customer review.

Simple Western lane view shows a specific band for Tyrosine Hydroxylase in 0.2 mg/mL of PC-12 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Western blot of 10 ug of rat striatal lysate showing specific immunolabeling of the ~60 kDa tyrosine hydroxylase protein.

10x Magnification cryo-sections mouse: primary antibody: anti-rb TH: 1:500; o.n. secondary antibody: 1:500 dk-anti-rb DyLight488. IHC-Fr image submitted by a verified customer review.

Tyrosine hydroxylase immunoreactivity in the mushroom body. All panels show a right brain hemisphere (dorsal to the top, medial to the right), except panels M-O (posterior to the top). Tyrosine hydroxylase (TH) immunoreactivity is shown in magenta (A, D, G, J, M) and synapsin immunoreactivity in green (B, E, H, K, N). Corresponding merged images in C, F, I, L, O. Scale bars = 100 um. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0160531), licensed under a CC-BY license.

Western blot analysis confirms activation of UPR following Hotel protein lysates from the right adrenal medulla of saline (2RS) and RH groups (2RH) were subjected to Western blot analyses as described in methods. Proteins were separated by SDS-PAGE, electroblotted and the membranes were sequentially probed with antibodies specific to Grp78, Derlin1, TH and GAPDH. Similar results were obtained in two separate replicate experiments. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0172789), licensed under a CC-BY license.

Impact of extrinsic and intrinsic hypoxia on catecholamine biosynthesis. Immunohistochemical staining showed an increased expression of Tyrosine Hydroxylase (red) in the necrotic and hypoxic core of the spheroid (DAPI, blue). Scale bar: 200 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31035382/) licensed under a CC-BY license.

Immunohistochemical analysis of a formalin fixed and paraffin embedded rat brain tissue section using Tyrosine Hydroxylase antibody at 1:5000 dilution. The primary antibody binding to its antigen was detected using HRP anti-Polyvalent ready-to-use kit (Ultratek, 35368) with DAB (brown) and the sections were further counterstained using hematoxylin. Isotype control section was incubated with Rabbit IgG Isotype Control antibody and was processed under the same assay conditions. Tyrosine hydroxylase staining (brown) was observed in the test samples only and the signal was specifically localized to the neuronal cells.

Immunostaining of whole-mount Drosophila brains using NB300-109 at 1:500 dilution. The tyrosine hydroxylase antibody worked really well and produced a bright staining with almost no background. Data courtesy of Dr. Olga Alekseenko, Neurobiology Dept. Harvard Medical School.

Immunohistochemical analysis of tyrosine hydroxylase in rat mesenteric artery-whole. ICC/IF image submitted by a verified customer review.

Western blot analysis confirms activation of UPR following RH.Total protein lysates from the right adrenal medulla of saline (2RS) and RH groups (2RH) were subjected to Western blot analyses as described in methods. Proteins were separated by SDS-PAGE, electroblotted and the membranes were sequentially probed with antibodies specific to Grp78, Derlin1, TH and GAPDH. Similar results were obtained in two separate replicate experiments.

Image showing differentiation of neurospheres as evidenced morphologically (A) and through staining by Tyrosine Hydroxylase (B) to show the ability to derive dopaminergic neurons from the rat progenitor cells.

Validation of protein alterations and functional studies for pathways induced by genetic and toxic stress, and reversal of PD-related alterations following S-HSP90 inhibition. c) Western blot and quantification of rotenone stress increase in the p-TH:TH ratio and its reduction by PU-H71 (200 nM). p-ERK/ERK, cell viability control; beta-actin, protein loading control. Mean +/- SEM, n = 3 individual values from the different experiments shown as points, One-way ANOVA with Tukey's post-hoc, ***p 10 larvae/sample). The Western blot was probed with antibodies against Lc3 and Actin as a loading control and is representative for three independent experimental repeats. The Lc3-II/Lc3-I ratio is indicated below. (F) Representative confocal micrographs of Ubiquitin antibody co-localization with Mm clusters in infected embryos/larvae at 1, 2 and 3 dpi. Scale bars, 10 μm. (G) Quantification of the percentage of Mm clusters positive for ubiquitin staining at 1 and 2 dpi (≥ 15 embryo/group). The results are accumulated from two independent experiments. ns, non-significant, *p10 larvae/sample). The Western blot was probed with an antibody detecting both poly- and mono-ubiquitin and with anti-Actin antibody as a loading control and is representative for three independent experimental repeats. (I) Representative confocal micrographs of GFP-Lc3 and Ubiquitin co-localization with Mm clusters in infected larvae at 3 dpi. Scale bars, 10 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30818338), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Ch, Dr, I, Ma

Species Glossary

Reactivity: Hu, Mu, Rt, Ch, Dr, I, Ma

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

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NB500-249

Beclin 1 Antibody - BSA Free

(9 Reviews)

(154 Publications)

0.1 ml

NB500-249

Immunohistochemistry: Beclin 1 Antibody - BSA Free [NB500-249] - Detection of Beclin 1 (red) in Pheochromocytes of the Adrenal Medulla 40x.

Flow Cytometry: Beclin 1 Antibody - BSA Free [NB500-249] - An intracellular stain was performed on U-87MG cells with Beclin 1 Antibody NB500-249 (blue) and a 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 2.5 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Western Blot: Beclin 1 Antibody - BSA Free [NB500-249] - Representative Western blot images and quantitative analyses of Beclin 1 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 = 12.37 for Beclin 1. *P 25 cells each. (E) GamGFP forms IR-induced foci in Ku80-defective MEFs. (F) Zoomed image from E. (G) IR-induced foci containing Gam only, 53BP1 only or both Gam and 53BP1 (>2600 total foci counted in three independent experiments). Error bars, SEM. Scale bars = 5 μm.DOI:
//dx.doi.org/10.7554/eLife.01222.012GamGFP marks DSBs in mammalian cells.(A) Live analysis of GamGFP localization to laser-induced DNA damage. Hela cells producing GamGFP were laser damaged along the cell track indicated by the red line at 0 min (m) and images were taken at the indicated times as shown. (B) GamGFP co-localizes with gamma H2AX in fixed Hela cells.DOI:
//dx.doi.org/10.7554/eLife.01222.013Ku inhibits GamGFP recruitment at DSBs independently of non-homologous end joining.Cells lacking either Ku80 or LigIV are defective in non-homologous end joining (NHEJ), yet the presence of Ku still inhibits recruitment of GamGFP to laser-induced DSBs even in NHEJ-defective cells, and thus independently of the cell's ability to complete NHEJ. Whereas it could have been possible that reduced GamGFP recruitment in the presence of Ku was caused by reduced persistence of DSBs due to their repair by NHEJ, our data show instead that Ku inhibits recruitment independently of successful NHEJ and imply that Ku binding to DSEs itself is inhibitory.DOI:
//dx.doi.org/10.7554/eLife.01222.014 Image collected and cropped by CiteAb from the following open publication (//elifesciences.org/articles/01222), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB500-249AF350
Alexa Fluor 405	NB500-249AF405
Alexa Fluor 488	NB500-249AF488
Alexa Fluor 532	NB500-249AF532
Alexa Fluor 594	NB500-249AF594
Alexa Fluor 647	NB500-249AF647
Alexa Fluor 700	NB500-249AF700
Alexa Fluor 750	NB500-249AF750
Allophycocyanin	NB500-249APC
Biotin	NB500-249B
DyLight 350	NB500-249UV
DyLight 405	NB500-249V
DyLight 488	NB500-249G
DyLight 550	NB500-249R
DyLight 594	NB500-249DL594
DyLight 650	NB500-249C
DyLight 680	NB500-249FR
DyLight 755	NB500-249IR
FITC	NB500-249F
HRP	NB500-249H
Janelia Fluor 549	NB500-249JF549
Janelia Fluor 646	NB500-249JF646
PE	NB500-249PE
PerCP	NB500-249PCP
mFluor Violet 450 SE	NB500-249MFV450
mFluor Violet 500 SE	NB500-249MFV500
mFluor Violet 610 SE	NB500-249MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB500-249SS

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NB100-56566

TLR4 Antibody (76B357.1) - BSA Free

(5 Reviews)

(138 Publications)

0.1 mg

NB100-56566

Immunohistochemistry: TLR4 Antibody (76B357.1) [NB100-56566] - Pericryptal Myofibroblasts are Responsible for Increased TLR4 Expression in a Subset of CRCs. Double-stained immunofluorescence for TLR4 (green) and vimentin (red) in normal (I), adenoma (II), and colon adenocarcinoma (III) (10x). In the stromal compartment of CRCs, immunofluorescent staining for TLR4 localized to the pericryptal myofibroblasts in a subset of samples. Image collected and cropped by CiteAb from the following publication (//www.jeccr.com/content/33/1/45), licensed under a CC-BY license.

Western Blot: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis using 2 ug/mL on (A) human intestine and 6 ug/mL on (B) mouse intestine and (C) rat intestine lysate.

Flow (Intracellular): TLR4 Antibody (76B357.1) [NB100-56566] - Analysis using PE conjugate of NBP2-27149. An intracellular stain was performed on Jurkat cells with TLR4 antibody (76B357.1) NBP2-27149PE (blue) and an isotype control MAB004 (orange). 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. Both antibodies were conjugated to phycoerythrin.

Immunocytochemistry/Immunofluorescence: TLR4 Antibody (76B357.1) [NB100-56566] - RH-30 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-TLR4 Antibody (76B357.1) NB100-56566 at 1 ug/ml overnight at 4C and detected with an anti-mouse 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.

Immunohistochemistry: TLR4 Antibody (76B357.1) [NB100-56566] - Immunofluorescent staining of TMAs. Representative tissue cores from normal (I), adenomatous polyps (II), and CRC (III and IV) are shown. Image collected and cropped by CiteAb from the following publication (//www.jeccr.com/content/33/1/45), licensed under a CC-BY license.

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis of rat salivary gland tissue section at 1:100 dilution. The antibody generated a membrane-cytoplasmic staining in the tissue with stronger signal in ductal epithelial cells.

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Tissue section of normal human skin stained with antibody at 5 ug/mL. Membrane-cytoplasmic immunopositivity of TLR4 was primarily observed in the pigmented basel cells and the adjacent keratinocytes in the epidermal layer.

Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - An intracellular stain was performed on RH30 cells with TLR4 Antibody (76B357.1) NB100-56566 (blue) and a matched mouse IgG2b Kappa isotype control (orange) MAB004. 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, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Human skin stained with at 5 ug/mL, peroxidase-conjugate and DAB chromogen. Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min.

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis of TLR4 in FFPE human colon tissue using an isotype control (top) and NB100-56566 (bottom) at 5 ug/mL.

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Human testis tissue stained with antibody at 5 ug/mL.

Immunohistochemistry-Frozen: TLR4 Antibody (76B357.1) [NB100-56566] - This image is TLR4(green) and nucleus(blue) at area postrema of the adult male mouse brain, x20 magnification. Primary antibody diluted 1:500. IHC-Fr image submitted by a verified customer review.

Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis of formaldehyde fixed THP-1 cells (human monocytic leukemia cells) using 2 ug/10^6 cells TLR4 antibody (clone 76B357.1) with detection employing a donkey anti-mouse IgG (H+L) cross adsorbed secondary antibody, (DyLight 488 conjugated). Isotype control samples incubated with mouse IgG2b isotype control antibody were processed in parallal under the same assay conditions.

Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - An intracellular stain was performed on Jurkat cells with TLR4 Antibody (76B357.1) NB100-56566 (blue) and a matched mouse IgG2b Kappa isotype control (orange) MAB004. 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, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).

Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - An intracellular stain was performed on Raw264.7 cells with TLR4 Antibody (76B357.1) NB100-56566 (blue) and a matched mouse IgG2b Kappa isotype control (orange) MAB004. 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, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).

Representative images of immunohistochemical staining of pancreatic adenocarcinoma upregulated factor (PAUF) and toll-like receptor 4 (TLR4) in human ovarian cancer specimens. The top row (A,B) represents normal ovarian tissue, middle row (C,D) shows immunoreactivity of ovarian carcinomas and the bottom row (E,F) indicates immunonegativity of ovarian caricinoma. The stromal cells of normal ovarian tissue show immunonegativity for PAUF and TLR4. The scale bar represents 100 μm.

ALWPs significantly reduced LPS-induced cell-surface levels of TLR4. (C) BV2 microglial cells were pretreated with ALWPs (500 ug/ml) or PBS for 30 min, followed by treatment with LPS (1 ug/ml) or PBS for 5.5 h. Cell-surface biotinylation was then conducted with a TLR4 antibody recognizing the N-terminal region of TLR4. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30319390), licensed under a CC-BY licence.

Effect of Cucumis sativus L. extract (CSE) on LPS induced TLR4 expression. b) Representative Western Blot of TLR4 and relative housekeeping alpha-tubulin were reported. c Expression of TLR4 protein in pAECs treated with LPS (10 ug/ml) in the presence or absence of increasing doses CSE (0.02; 0.2; 2 mg/ml); data shown are representative of at least three independent experiments and represent the mean +/- SEM. Different letters above the bars indicate significant differences (p < 0.05 ANOVA post hoc Tukey's test) Image collected and cropped by CiteAb from the following publication (//bmccomplementalternmed.biomedcentral.com/articles/10.1186/s12906-018-2254-1), licensed under a CC-BY licence.

Dasatinib affects TLR4 signaling to regulate LPS-induced proinflammatory cytokine levels in BV2 microglial cells. d) BV2 microglial cells were treated with dasatinib (250 nM) or vehicle (1% DMSO) for 30 min followed by LPS (1 ug/ml) or PBS for 5.5 h, and cell surface biotinylation was conducted. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31655606), licensed under a CC-BY licence.

Corroboration of human AKR2–protein interactions with highest confidence. (A) Representation of the components of the protein pull-down experiment using c-Myc magnetic beads with Myc-tagged human AKR2. (B) Western blot analysis of AKR2–protein interactions. Interacting proteins were incubated with AKR2 and immunopresipitated with c-Myc magnetic beads specific for the AKR2 protein tag (I) or with the c-Myc magnetic beads only as negative control (C-). Recombinant interacting proteins were included as positive control (C+). Mouse or rabbit antibodies specific for each protein were used as primary antibodies and then anti-mouse or anti-rabbit secondary antibodies were used to identify the presence of the interacting proteins (red arrow). The origin of the primary antibody is shown. (C) Corroboration of human AKR2–AKR2 interaction using protein pull-down with c-Myc magnetic beads with human Myc-tagged AKR2. The experiment was conducted as described in (B). The origin of the primary antibody and predicted size for AKR2 monomer, dimer, and trimer together with bands corresponding to fragments of the mouse anti-c-Myc antibody attached to magnetic beads are shown. (D) In order to identify unrelated proteins eluted from the c-Myc magnetic beads, a similar experiment was conducted with selected proteins but incubating only with the secondary antibody. A lane of SDS-PAGE stained with Bio-Safe Coomassie Stain corresponding to c-Myc magnetic beads alone eluted using Laemmli sample buffer was included (C--). The bands corresponding to fragments of the mouse anti-c-Myc antibody attached to magnetic beads are shown. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32046307), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$TDP1 depletion sensitizes glioblastoma-resistant cancer cells to temozolomide. (a) Cell lysates from RKO and DLD1 cancer cell lines were fractionated by SDS–PAGE and analyzed by immunoblotting using anti-TDP1 (Eurogentec), anti-Top1 (Santa Cruz), anti-Tubulin (Abcam) and anti-APE1 (Novus) antibodies (left). RKO and DLD1 cells were compared for their survival following exposure with the indicated doses of MMS for 15 min at 37°C (right). Data are the average of three independent experiments ± s.e.m. Where not visible, error bars are smaller than the symbol. (b) RKO cells were subjected to scrambled siRNA (Mock) or siRNA against TDP1 (TDP1KD) and cell lysates analyzed by immunoblotting (left). Control RKO (RKOSc siRNA) and RKO cells in which TDP1 levels were depleted (RKOTDP1 siRNA) were examined for their survival following exposure to the indicated doses of MMS, as described earlier in the text (right). (c) Glioblastoma multiforme T98G and U87 cancer cell lines were analyzed for methylguanine methyltransferase (MGMT) expression using anti-MGMT antibodies (Abcam) (left). High-MGMT expressing T98G cells were compared with low-MGMT expressing U87 cells for their survival following exposure to increasing concentrations of the alkylating agent temozolomide (right). (d) TG98 cells were subjected to scrambled siRNA (Mock) or siRNA against TDP1 (TDP1KD) and cell lysates analyzed by immunoblotting (left). Control T98G cells (T98GSc siRNA) and cells in which TDP1 levels were depleted (TG98TDP1 siRNA) were incubated with DMSO or 150 μM of the APE1 inhibitor CRT0044876 ‘APEi’ for 2 h followed by additional incubation with the indicated concentrations of MMS for 15 min at 37°C. Cell survival was blindly scored from three independent biological repeats, and data are the average ± s.e.m. (right). (e) Model for the repair of temozolomide-induced DNA breaks. Temozolomide delivers a methyl group to purine bases of DNA, resulting in O6-methyl guanine, N7-methyl guanine and N3-methyl adenine. The primary cytotoxic lesion is believed to be O6-methyl guanine, which can be removed by a direct repair mechanism mediated by methylguanine methyltransferase (MGMT). Inherent and acquired resistance to temozolomide via MGMT expression presents a major challenge in cancer therapy, particularly for glioblastoma multiforme. TDP1 promotes the repair of methylated purines induced by temozolomide via a distinct non-canonical BER pathway. Increasing the load of unrepaired methylated purines by exploiting the limited availability of TDP1 alone or in combination with canonical BER factors such as APE1 provides a new synthetic lethal setting to improve the clinical outcome of temozolomide-based cancer therapy. Asterisks denote statistical differences (P < 0.05; t-test) between control and TDP1-deficient cells. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/24335147), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

OTULIN overexpression reduced the number of Iba-1+ microglial cells in the cortical ischemic penumbra in focal cerebral ischemia/reperfusion rats. A Representative images of Iba-1+ microglial cells that were morphologically classified into three types; scale bar = 10 μm. B Iba-1+ microglial cells in the ischemic penumbra and contralateral cortex of each group were detected by confocal immunofluorescence at 24 h, 72 h, and 7 days; scale bar = 75 μm. C The histogram represents the quantitative analysis of mean Iba-1 immunofluorescence intensity in each group (n = 6). Data are presented as the means ± SEMs; ###P < 0.001 versus the tMCAO group; &&&P < 0.001 versus the tMCAO+LV-Scramble group Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29544517), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

FKBP51 modulates EMT hallmarks. (a and b) FKBP51 silencing of SAN melanoma cells decreases TGF-beta and T beta RIII expression. Normalized expression rates (mean±S.D.) of TGF-beta (a, left) and T beta RIII (b, left) mRNA levels; (a, n=3; b, n=5). FKBP51 siRNA-treated sample expression=1. Western blot assay of TGF-beta in SAN melanoma cells treated with nonsilencing (NS) RNA or FKBP51 siRNA (a, right). Flow cytometric analysis of T beta RIII (endoglin) expression (b, right): FKBP51 siRNA-treated cells moved to the left relative to NS RNA-treated cells because of reduced staining (MFI: 111±10 and 64±3 in NS RNA and FKBP51 siRNA-treated cells, respectively). (c, and d) FKBP51 overexpression enhances TGF-beta and T beta RIII expression. Normalized expression of TGF-beta (c, left) and T beta RIII (d, left) mRNA in WT, EV-, or FKBP51-stably transfected melanoma cells. WT sample expression=1. (c, n=3; d, n=5). Western blot assay of TGF-beta levels in the same cells (c, right). Anti-Flag-labelled exogenous FKBP51. Flow cytometric analysis of T beta RIII expression (d, right). FKBP51-overexpressing cells moved to the right relative to WT or EV cells, indicating increased expression. (e) Graphic representation of cell counts ( × 10−3) in WT, EV-, or FKBP51-stably transfected melanoma cell cultures. Results are means of six independent experiments each performed in triplicate. (f) FKBP51 upregulates colony number and size. Colonies were counted under low magnification ( × 10) at four points on each dish. The data represent the means of two independent experiments performed in triplicate. Magnification: upper, × 10; lower × 40. (g) Boyden chamber filters invaded by melanoma cells. Filter was stained with Crystal Violet. OD value was obtained by reading the eluted Crystal Violet with a spectrophotometer at a wavelength of 570 nm. The increased OD in FKBP51 samples indicated that FKBP51-overexpressing cells invaded matrigel more efficiently than control cells. The data represent the means of three independent experiments performed in triplicate Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/cddis2013109), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Functional assessment of Fanconi anemia pathway. (a) The experimental scheme for MMC treatment. Twenty‐four hours after plating, cells were cultured with or without MMC 1 μM for an additional 24 hr, after which the cells were harvested for western blot or immunostaining. (b) Western blot with FANCD2 antibody of BJ, proband fibroblasts, FANCB mutant (null) fibroblasts and FANCD2 mutant (null) fibroblasts. (c) Western blot with FANCD2 antibody of non‐FA control lymphoblasts (LCL), proband LCL‐A‐2017, proband LCL‐B‐2017, FANCB mutant (null) LCL, and FANCD2 mutant (null) LCL. (d) Representative figures of FANCD2 foci formation in the indicated cells. (e) Quantification of FANCD2 foci formation following treatment with or without 1 μM MMC. Experiments were performed in triplicate. One hundred cells were counted for each experiment Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29193904), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Pot1a prevents DDR in HSCs. a, b Telomeric DDR in 8 week-old LSK cells upon Pot1a knockdown. a Immunocytochemical staining of TRF1 (green) and 53BP1 (red). Foci co-stained with TRF1 and 53BP1 were identified as TIFs. Nuclei were stained with TOTO3 (blue). Scale bar, 2 μm. b Frequency of TIFs per cell after 1 week of culture. Data are expressed as the mean ± SD (n = 100, *p < 0.01 by t-test). Representative data from three independent experiments are shown. c–f Telomeric DDR in donor-derived control-GFP or Pot1a overexpressing 8 week-old LSK cells 4 months post BMT. c Immunocytochemical staining of TRF1 (green) and 53BP1 (red). Nuclei were stained with TOTO3 (blue). Scale bar, 2 μm. d Frequency of TIFs per cell. Data are expressed as the mean ± SD (n = 120, *p < 0.01 by t-test). Representative data from 3 independent experiments are shown. e Immunocytochemical staining of TRF1 (green) and RPA32 (red). f Immunocytochemical staining of TRF1 (green) and pChk1 (red). Scale bar, 2 μm. g Flow cytometric analysis of Chk1 and pChk1 in donor-derived GFP+ LSKCD48–CD150+ cells after 5 months of 2nd BMT. Mean fluorescence intensity of Chk1 and pChk1 (left panels). Data are expressed as the mean ± SD (n = 3, *p < 0.01 by t-test). Representative FACS profiles of Chk1 and pChk1 in donor-derived GFP+ LSKCD48–CD150+ cells (right panels) Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28986560), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Lamin A/C-53BP1 interaction is regulated in a DNA damage-dependent manner. (A) Left; HDF were subjected to IR (10 Gy) and allowed to recover for 1 h. Association between A-type lamins and 53BP1 was assessed by immunoprecipitation of endogenous lamin A/C followed by immunoblotting with 53BP1 and lamin A/C antibodies. WCE, whole cell extract, IP: immunoprecipitates. WCE represents 5% input. Right; endogenous lamin A/C was immunoprecipitated and supernatants or pellets were analysed by immunoblotting for interaction with JMJD2A. (B) U2OS/GFP-lamin A cells were pretreated with caffeine (20 mm) for 1 h before exposure to IR (10 Gy, 1 h recovery). Cell extracts were then subjected to immunoprecipitation using GFP-Trap beads, and bound complexes were then analysed by immunoblotting using 53BP1 and GFP antibodies. WCE represents 1% input. (C) As in (C) except cells were pretreated with 10 μm ATMi for 1 h before IR. WCE represents 1% input. (D) U2OS/GFP-lamin A cells were subjected to laser micro-irradiation, fixed 1 h later and immunostained with gamma -H2AX antibody. Scale bar, 10 μm. (E) U2OS cells were transfected with siCTRL or siLMNA and subjected to laser micro-irradiation, fixed 15 min later and then processed for immunofluorescence with gamma -H2AX and 53BP1 antibodies. Scale bar, 10 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/25645366), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Bv, Ma, Pm, ChHa

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Ma, Pm, ChHa

Applications:

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

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

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Alt Catalog#

IMG-5031A

Alt Catalog# IMG-5031A

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP2-27149AF350
Alexa Fluor 405	NBP2-27149AF405
Alexa Fluor 488	NBP2-27149AF488
Alexa Fluor 532	NBP2-27149AF532
Alexa Fluor 594	NBP2-27149AF594
Alexa Fluor 647	NBP2-27149AF647
Alexa Fluor 700	NBP2-27149AF700
Alexa Fluor 750	NBP2-27149AF750
Allophycocyanin	NBP2-27149APC
Allophycocyanin/Cy7	NB100-56566APCCY7
Biotin	NBP2-27149B
DyLight 350	NBP2-27149UV
DyLight 405	NBP2-27149V
DyLight 488	NBP2-27149G
DyLight 550	NBP2-27149R
DyLight 594	NB100-56566DL594
DyLight 650	NBP2-27149C
DyLight 680	NBP2-27149FR
DyLight 755	NBP2-27149IR
FITC	NBP2-27149F
HRP	NBP2-27149H
Janelia Fluor 549	NB100-56566JF549
Janelia Fluor 646	NB100-56566JF646
PE	NBP2-27149PE
PE/Atto594	NB100-56566PEATT594
PE/Cy5.5	NB100-56566PECY55
PE/Cy7	NB100-56566PECY7
PerCP	NBP2-27149PCP
mFluor Violet 450 SE	NBP2-27149MFV450
mFluor Violet 500 SE	NBP2-27149MFV500
mFluor Violet 610 SE	NBP2-27149MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Azide Free	NBP2-27149
BSA Free	NB100-56566SS
Azide Free	NBP2-27149SS

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NB100-106

OGG1 Antibody - BSA Free

(5 Reviews)

(128 Publications)

0.1 ml

NB100-106

Immunohistochemistry: OGG1 Antibody [NB100-106] - Figure 5 IHC-P analysis of hOGG1 protein expression in serous ovarian cancer: (A) Normal ovarian tissue treated with anti-hOGG1 antibody, biotinylated anti-rabbit IgG secondary antibody, and avidin-biotin-peroxidase complex, and counterstained with p-dimethylaminobenzaldehyde reagent (magnification 100 x); (B) serous cystadenoma with positive staining (magnification 200 x); (C) LG-SOC tissue with moderate positive staining (magnification 200 x); and (D) HG-SOC with negative staining (magnification 200 x). Black arrow pointed at the immunostained epithelial cells. Image collected and cropped by CiteAb from the following publication (//ovarianresearch.biomedcentral.com/articles/10.1186/1757-2215-6-74), licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: OGG1 Antibody [NB100-106] - A431 cells were 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-hOgg1 [NB100-106] 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.

Immunohistochemistry-Frozen: OGG1 Antibody [NB100-106] - Detection of OGG1 in formaldehyde-fixed frozen sections of the substantia nigra from a Rhesus macaque (Macaca mulatta) using NB100-106 at 10 ug/mL. Photo courtesy of Glen Kisby, Oregon Health Sciences University.

Flow (Intracellular): OGG1 Antibody [NB100-106] - An intracellular stain was performed on HeLa cells with NB100-106C (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 Dylight 650.

Immunohistochemistry: OGG1 Antibody [NB100-106] - The H&E staining ((a)-(c)) and IHC staining for oxidative DNA damage, 8-oxo-dGuo ((d)-(g)), and DNA repair enzyme hOGG1 ((h)-(j)) in control subjects and tumor and nontumor lesions of BCC patients. Values given are mean +/- SD. The statistical significance of differences between the control and case and between adjacent epidermis and tumor lesions of BCC patients was evaluated by nonparametric variables with Kruskal-Wallis test followed by Dunnett's post hoc test. * P < 0.05, *P < 0.01, *P < 0.001 compared to control; ##P < 0.01, ###P < 0.001 compared to tumor lesions of BCC patients. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/omcl/2016/5934024/), licensed under a CC-BY license.

Western Blot: OGG1 Antibody [NB100-106] - The protein expression levels of HVL and LVL sorted cells by western blotting. The ATM, phospho-ATM, GPX2, MRE11, OGG1, and XPC levels were measured by western blot analysis. The figure is representative of three independent experiments. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0164281), licensed under a CC-BY license.

Immunohistochemistry-Paraffin: OGG1 Antibody [NB100-106] - Comparison of the expression of OGG1 protein in normal colon tissue, polyp and cancer tissue of adenoma (AD, n = 68) and carcinoma (CRC, n = 103) patients. Immunohistochemical detection in paraffin embedded sections stained with hematoxylin and eosin. Center mark in the box indicates the medians of the samples. The length of each box (IQR, interquartile range) represents the range within which the central 50% of the values fell, with the vertical edges placed at the first and third quartiles. Whiskers show variability outside the upper and lower quartiles. P was obtained with the Mann-Whitney test. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0115558), licensed under a CC-BY license.

Flow Cytometry: OGG1 Antibody [NB100-106] - An intracellular stain was performed on HeLa cells with OGG1 Antibody NB100-106 (blue) and a 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).

Immunohistochemistry: OGG1 Antibody [NB100-106] - Staining of human tonsil, germinal center and mantle zone.

Flow Cytometry: OGG1 Antibody [NB100-106] - Baseline Ogg1 expression in human PBMC from a healthy donor. Image from verified.customer review.

Flow Cytometry: OGG1 Antibody [NB100-106] - An intracellular stain was performed on Jurkat Cells with NB100-106 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, Dylight 550.

The protein expression levels of HVL and LVL sorted cells by western blotting.(A) The ATM, phospho-ATM, GPX2, MRE11, OGG1, and XPC levels were measured by western blot analysis. The figure is representative of three independent experiments. (B) The relative fold activity was quantified using AlphaImage software and normalized to the expression of the internal actin control. *P < 0.05, **P < 0.01.

HDAC1 modulates OGG1 activity.a, b) Representative images and quantification of OGG1 acetylation assessed by western blot analysis using antibody against acetylated lysine (Ac-Lysine, 4G12). Recombinant OGG1 (100 ng) was first acetylated by p300 (25 ng), and then incubated with titrated amount of HDAC1 (25 and 50 ng). a, b Four replicates from four independent experiments. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32424276), licensed under a CC-BY licence.

Comparison of the expression of OGG1 (A) and PARP-1 (B) protein in normal colon tissue, polyp and cancer tissue of adenoma (AD, n = 68) and carcinoma (CRC, n = 103) patients.Immunohistochemical detection in paraffin embedded sections stained with hematoxylin and eosin. Center mark in the box indicates the medians of the samples. The length of each box (IQR, interquartile range) represents the range within which the central 50% of the values fell, with the vertical edges placed at the first and third quartiles. Whiskers show variability outside the upper and lower quartiles. P was obtained with the Mann-Whitney test. Representative examples of the levels of PARP-1 protein in tissues of CRC patients determined by Western analysis. The analysis was performed on tumor and normal tissues of 41 CRC patients (C). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25526641), licensed under a CC-BY licence.

The H&E staining ((a)-(c)) and IHC staining for oxidative DNA damage, 8-oxo-dGuo ((d)-(g)), DNA repair enzyme, hOGG1 ((h)-(k)), and antioxidant proteins, CAT ((l)-(o)), GCLC ((p)-(s)), GPx ((t)-(w)), Nrf2 ((x)-(aa)), and MnSOD ((ab)-(ae)), in control subjects and tumor and nontumor lesions of BCC patients. Values given are mean +/- SD. The statistical significance of differences between the control and case and between adjacent epidermis and tumor lesions of BCC patients was evaluated by nonparametric variables with Kruskal-Wallis test followed by Dunnett's post hoc test. *P < 0.05, **P < 0.01, ***P < 0.001 compared to control; ##P < 0.01, ###P < 0.001 compared to tumor lesions of BCC patients. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27057281), licensed under a CC-BY licence.

Immunoblot analysis of kidney lysates.(A) Blot membranes were incubated with the antibodies against Nrf2, p53 and 8-oxoguanine glycosylase alpha (OGG1alpha). The mean integrated optical density is related to actin for Nrf2 (B), p53 (C) and mature 8-oxoguanine glycosylase alpha (D) levels. Results are given as a mean +/- S.E.M. KD4, KD6, KD8 - groups of Eker rats (Tsc2+/-) treated with HFKD for four, six or eight mo., respectively; ST - Eker rats fed with a standard diet; LE ST - wild-type Long Evans rats treated with a standard diet; LE KD - wild-type Long Evans rats treated with a ketogenic diet similarly to the KD6 group. *P < 0.05, **p < 0.01, ***p < 0.001 as compared to ST unless otherwise stated (horizontal line) for ANOVA with a Fisher post hoc test. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26892894), licensed under a CC-BY licence.

Immunohistochemistry of putative iron importers in chronic kidney disease. Representative images of ZIP8 (a–f), ZIP14 (g–l), and divalent metal transporter 1 (DMT1; m–r) staining in control (a,g,m), early diabetic nephropathy (DNE; b,h,n), advanced diabetic nephropathy (DNA; c,i,o), focal segmental glomerulosclerosis (FSGS; d,j,p), lupus nephritis (LN; e,k,q) and IgA nephropathy (IgAN; f,l,r). Intensity in proximal and distal tubules quantified (s). Dots represent all quantified images (5 images per biopsy). Renal structures indicated as glomerulus (G), proximal tubule (PT), distal tubule (DT). Scale bar 40 µM. **p < 0.01; ***p < 0.001. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29921869), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Cx43-knockdown cells regulate the expression of HIF-1 alpha , c-Src, VEGF and the proliferation of endothelial cells. (a) The expression of Cx43, c-Src, and HIF-1 alpha was measured in Cx43-knocknown cells by Western blot analysis. beta -actin expression served as loading controls for and total protein. Inserted values indicated relative protein expression in comparison with beta -actin; (b) Cx43-knockdown cells were cotransfected with pCLNCX-6× HRELuc and pTCYLacZ plasmids. Then, cells were treated with 100 ng/μL PP2 or DMSO for 1 h. At 6 h post-transfection, their luciferase activities were determined and normalized with beta -gal activity. Data shown were the mean ± SD (n = 4); (c) The conditioned medium of Cx43-knockdown cells was measured by ELISA. Data shown were the mean ± SD (n = 4); (d) The conditioned medium of Cx43-knockdown cells reduced the proliferation of endothelial cells. The HEMC-1 cells treated with conditioned medium of Cx43-knockdown cells were examined for proliferation activity. Cell viability was assessed by the WST-1 assay. Data shown were the mean ± SD (n = 4). * p < 0.05; ** p < 0.01; *** p < 0.001. Image collected and cropped by CiteAb from the following open publication (//www.mdpi.com/1422-0067/16/1/439), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

PHD3 inhibition reduces the amount of hyperphosphorylated Rb and increases p27 in hypoxia.(A) SCC2 cells were transfected with the indicated siRNAs and exposed to normoxia or hypoxia for 24 to 48 hours followed by western blot analysis of PHD3, phosphorylated Rb (p-Rb) and cyclin B1. (B) HeLa cells were transfected with the indicated siRNAs, synchronized and exposed to normoxia or hypoxia for 6 to 24 hours after release. PHD3, phosphorylated Rb (p-Rb) and cyclin D1 were analyzed from samples by western blotting. (C) SCC2 cells were transfected with the indicated siRNAs and exposed to normoxia or hypoxia for 24 hours followed by western blot analysis of p21(Cip1). (D) Cells transfected with the indicated siRNAs and exposed to normoxia or hypoxia for 24 hours followed by western blot analysis of p16 and p27. Hypoxia was monitored by HIF-1a expression. (E) Cells transfected with the indicated siRNAs and exposed to normoxia or hypoxia for 24 and 48 hours followed by western blot analysis of p27. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/22087251), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Lactating mammary gland.Smaller panels display enlargements of the indicated areas. Size bars: 50 μm, 40x obj was used for all micrographs. A. Collagen I IHC allows visualisation of the basement membrane surrounding the dilated ducts. B. HIF-1 alpha was not detected in the epithelial cells of the lactating gland (compare with Fig 1). C. Macrophage infiltration was sparse in the lactating mammary gland as judged by F4/80 IHC. D. A subset of cuboidal luminal epithelial cells was distinctively positive for HIF-2 alpha . E. The percentage of HIF-2 alpha -positive out of total luminal epithelial cells was counted in sections from three mice. Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.pone.0125771), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Expression of RPE-specific marker proteins in hPSC-RPE and iPSC-RPE cells.(a–b) Immunoblotting showing the expression of RPE-specific proteins BEST1, RPE65, CRALBP, and the loading control beta -Actin in hPSC-RPE (a) and iPSC-RPE (b) cells. Two gels/blots in the same panel were prepared from the same cell lysate of each PSC-RPE to detect BEST1 + beta -Actin, and RPE65 + CRALBP, respectively. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34061021), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Genetic or pharmacologic disruption of Caspase 1 blocks LukAB-induced cytokine secretion but not cell death.(A) THP1 cells were transfected with siRNA against Caspase 1, ASC or a non-targeting sequence. Cell lysates were collected and analyzed by immunoblot to confirm knockdown of pro-Caspase 1 and ASC. (B) THP1 siRNA cells were incubated with propidium iodide and intoxicated with 50 ng/mL LukAB for 1 hour then analyzed by flow cytometry. (C and D) THP1 siRNA cells were either primed with LTA (500ng/mL) (C) or untreated (D) then intoxicated with LukAB (50ng/mL) for 1 hr before culture supernatants were analyzed for release of IL-1 beta (C) or IL-18 (D). (E and F) Primary CD14+ human monocytes were incubated with the indicated concentration of z-YVAD-FMK or VX-765 for 30 minutes. Primary monocytes were incubated in the presence of propidium iodide (E) then intoxicated with LukAB (50ng/mL) and analyzed by flow cytometry. (F) Primary monocytes were intoxicated with LukAB (50ng/mL) and culture supernatants were analyzed for release of IL-18. Propidium iodide staining and IL-18 secretion are reported as a fraction of measurement in primary cells not treated with inhibitor. Bars represent the mean ± standard error of the mean for at least two independent experiments, each performed in triplicate. Asterisks indicate significance at a p-value of ≤ 0.05 by Tukey’s multiple comparisons post-test for 1-way or 2-way ANOVA, as appropriate. Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.ppat.1004970), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Reactivity:

Hu, Mu, Rt, Po, Pm, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Pm, Rb

Applications:

, , , , , , ,

Applications: , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-106AF350
Alexa Fluor 405	NB100-106AF405
Alexa Fluor 488	NB100-106AF488
Alexa Fluor 532	NB100-106AF532
Alexa Fluor 594	NB100-106AF594
Alexa Fluor 647	NB100-106AF647
Alexa Fluor 700	NB100-106AF700
Alexa Fluor 750	NB100-106AF750
Allophycocyanin	NB100-106APC
Allophycocyanin/Cy7	NB100-106APCCY7
Biotin	NB100-106B
DyLight 350	NB100-106UV
DyLight 405	NB100-106V
DyLight 488	NB100-106G
DyLight 550	NB100-106R
DyLight 594	NB100-106DL594
DyLight 650	NB100-106C
DyLight 680	NB100-106FR
DyLight 755	NB100-106IR
FITC	NB100-106F
HRP	NB100-106H
Janelia Fluor 549	NB100-106JF549
Janelia Fluor 646	NB100-106JF646
PE	NB100-106PE
PE/Atto594	NB100-106PEATT594
PE/Cy5.5	NB100-106PECY55
PE/Cy7	NB100-106PECY7
PerCP	NB100-106PCP
mFluor Violet 450 SE	NB100-106MFV450
mFluor Violet 500 SE	NB100-106MFV500
mFluor Violet 610 SE	NB100-106MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-106SS

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NB300-144

Laminin Antibody

(9 Reviews)

(122 Publications)

0.1 ml

NB300-144

MxIF analysis of mouse submandibular salivary gland morphogenesis. MxIF of a developmental TMA including embryonic stages (E14, E16, E18) and postnatal stages (P5 and P20) was performed using sequential application of directly conjugated antibodies to detect multiple markers of tissue structures and cell types on the same tissue sections. Tissue compartments. The epithelium, mesenchyme, neurons, and basement membranes was detected using antibodies directed towards E-cadherin (ECAD, red), platelet-derived growth factor (PDGFR, green), beta III tubulin (bIII, magenta), and Laminin Antibody [NB300-144] (LMN, cyan), respectively. Image collected and cropped by CiteAb from the following publication (//bio.biologists.org/cgi/doi/10.1242/bio.20134309), licensed under a CC-BY license.

Staining of mouse section of cortex stained with Laminin Antibody [NB300-144] (red). Blue is DAPI staining of DNA. This antibody reveals strong staining in the basement membranes of blood vessels.

Staining of rat spinal cord and dorsal root paraformaldehyde/paraffin-embedded tissue using Laminin Antibody [NB300-144]. Pepsin antigen retrieval was performed on this tissue sample.

Analysis of rat heart cells lysates (lane 1) and 0.2 ug of purified laminin protein from mouse EHS sarcoma (lane 2) Laminin Antibody [NB300-144] recognizes 3 laminin isotypes: alpha 1 (440 kDa), beta 1 (220 kDa) and gamma 1 (220 kDa). Also recognized is a laminin binding protein at 120 kDa in both rat heart lysates and purified laminin protein. Since this protein always coexpresses with laminin this crossreactivity is irrelevant. Theoretical molecular weight of LAMA1 is 337 kDa.

IF Confocal analysis of HeLa cells using Laminin Antibody [NB300-144] (1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green, A). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red, B). DAPI was used to stain the cell nuclei (blue, C).

Western blot analysis of the effects of IP6, Ins, IP6 + Ins and normal saline on the levels of collagen IV, Laminin and Fibronectin, with Laminin Antibody [NB300-144]. IP6 or Ins treatment decreased the protein expression of LN and FN, and the combined IP6 + Ins treatment resulted in significantly greater effects compared with treatment with either compound alone. The Western blot membranes were stripped and reprobed for beta-actin as an internal control to confirm equal loading. (A) representative blots from one of three separate experiments; (B) relative band intensities based on densitometry. The results are expressed as the mean +/- standard deviation from three independent experiments. * p < 0.05 compared to the IP6 + Ins group; #p < 0.05 compared to the normal saline group. Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/2072-6643/8/5/286), licensed under a CC-BY license.

Immunofluorescence staining for extracellular matrix proteins in day 1 and day 7 HWJSC spheroids. HWJSC spheroids were cultured for either 1 day or 7 days (with 6 days of osteogenic differentiation), fixed, and stained for extracellular matrix protein: laminin (C) and counter-staining with Hoechst. Scale bars represent 100 um. Image collected and cropped by CiteAb from the following publication (journals.plos.org/plosone/article?id=10.1371/journal.pone.0184155), licensed under a CC-BY license. Using the Alexa Fluor 488 format of this antibody.

Laminin staining on an E12.5 mouse Right Ventricle. IHC-Fr image submitted by a verified customer review.

A surface stain was performed on HeLa cells with the Alexa Fluor 647 conjugate of Laminin Antibody [NB300-144AF647] (blue) and a matched isotype control (orange). Cells were incubated in an antibody dilution of 2.5 ug/mL for 20 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Analysis of Laminin-1 expression in mouse EHS tumor crude extracts (left) and Laminin-2 expression in rat heart crude extracts (right), using Laminin Antibody [NB300-144]. The Laminin polyclonal antibody was used at 1 ug/mL. Theoretical Molecular Weight of NB300-144 is 337 kDa.

VSOP observed in perivascular-restricted spinal cord lesions with intact BBB. Immunostaining for laminin (brown) using Laminin Antibody [NB300-144] shows vascular endothelium and glia limitans of a perivascular lesion, along with infiltrating cells and VSOP (blue). Image collected and cropped by CiteAb from the following publication (//asn.sagepub.com/lookup/doi/10.1042/AN20120081), licensed under a CC-BY license.

SOX17 regulates adult muscle regeneration after injury in Pax7CreERT2/+;Sox17fl/fl mutant mice. Representative images of cryosections from regenerating adult TA muscles d28 after injury, showing immunofluorescence for PAX7+ (quiescent, arrows) cells. Scale bar, 25 Aum. Image collected and cropped by Citeab from the following publication (SOXF factors regulate murine satellite cell self-renewal and function through inhibition ofI2-catenin activity. Elife (2018)) licensed under a CC-BY license.

VSOP observed in perivascular-restricted spinal cord lesions with intact BBB (B) Immunostaining for laminin (brown) shows vascular endothelium and glia limitans of a perivascular lesion, along with infiltrating cells and VSOP (blue). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23452162), licensed under a CC-BY licence.

Reactivity:

Hu, Mu, Rt, ChHa, In, Ma, Rb, Sh

Species Glossary

Reactivity: Hu, Mu, Rt, ChHa, In, Ma, Rb, Sh

Applications:

, , , ,

Applications: , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB300-144AF350
Alexa Fluor 405	NB300-144AF405
Alexa Fluor 488	NB300-144AF488
Alexa Fluor 532	NB300-144AF532
Alexa Fluor 594	NB300-144AF594
Alexa Fluor 647	NB300-144AF647
Alexa Fluor 700	NB300-144AF700
Alexa Fluor 750	NB300-144AF750
Allophycocyanin	NB300-144APC
Allophycocyanin/Cy7	NB300-144APCCY7
Biotin	NB300-144B
DyLight 350	NB300-144UV
DyLight 405	NB300-144V
DyLight 488	NB300-144G
DyLight 550	NB300-144R
DyLight 594	NB300-144DL594
DyLight 650	NB300-144C
DyLight 680	NB300-144FR
DyLight 755	NB300-144IR
FITC	NB300-144F
HRP	NB300-144H
Janelia Fluor 549	NB300-144JF549
Janelia Fluor 646	NB300-144JF646
PE	NB300-144PE
PE/Atto594	NB300-144PEATT594
PE/Cy5.5	NB300-144PECY55
PE/Cy7	NB300-144PECY7
PerCP	NB300-144PCP
mFluor Violet 450 SE	NB300-144MFV450
mFluor Violet 500 SE	NB300-144MFV500
mFluor Violet 610 SE	NB300-144MFV610

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NB100-2284

CD31/PECAM-1 Antibody

(6 Reviews)

(122 Publications)

0.1 ml

NB100-2284

Staining of CD31/PECAM-1 in Rat Liver sample. IHC-P image submitted by a verified customer review.

Human lung adenocarcinoma using. Antibody at 1:100 with citrate epitope retrieval at pH 6.0.

Analysis in rat muscle blood vessels using. IHC-P image submitted by a verifed customer review.

Matrix-HA supplemented with strontium was implanted subcutaneously in mice, where immunochemistry analysis and histology occurred of the newly formed tissues. (C) Within each matrix: Matrix-HA, Matrix-8Sr-HA and Matrix-50Sr-HA, were indicated by immunostaining of CD31 for the newly formed tissues. (D) Number of vessels within each tissue was quantified using NDP view software. Unit is square mm. Immunostaining analysis of slides occurred for 2 samples per condition and 3 sections were analyzed per sample and per group of matrix. Citation: Ehret C, Aid-Launais R, Sagardoy T, Siadous R, Bareille R, Rey S, et al. (2017) Strontium-doped hydroxyapatite polysaccharide materials effect on ectopic bone formation. PLoS ONE 12(9): e0184663. //doi.org/10.1371/journal.pone.0184663

Imaging of HUVEC monolayer on glass substrate. Fixation with 4% PFA. Blocking with Goat Serum at 1:100. Incubation 2 hours at room temperature. ICC/IF image submitted by a verified customer review.

Analysis of canine kidney tissue using CD31/PECAM-1 antibody. Antibody was used at a 1:50 concentration in Casein in PBS and left at 4C overnight on paraffin embedded canine bladder tissue. HIER was performed in Tris/EDTA buffer, pH 9 for two hours at 75C. Image from verified customer review.

Analysis in pig SC cells using CD31/PECAM-1 Antibody (red). Blue color showing nucleus labeling. ICC/IF image submitted by a verified customer review.

Immunohistochemistry evaluation of CD31/PECAM-1 expression. Representative images of CD31/PECAM-1 staining of vessels in the three groups of implanted matrices (scale bar = 100 um). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-06258-0) licensed under a CC-BY license.

CD31 staining on E11.5 mouse Pharyngeal Mesoderm. Fixed with 4% PFA overnight. Blocked with 1% BSA. Primary antibody at 1:100. Secondary antibody at 1:1000, conjugated to Alexa Fluor 488. IHC-Fr image submitted by a verfied customer review.

Immunohistochemistry evaluation of CD31 expression. (a) Representative images of CD31 staining of vessels in the three groups of implanted matrices (scale bar = 100 µm). (b) Quantification of vessel density inside the implanted matrices at 5 weeks post implantation (n = 6; Average ± SD). NS and **denote Non Significant and p < 0.01, respectively.

Impact of T-AuNPs on endothelial cells in vivo. Immunofluorescent staining for endothelial cells using an anti-CD 31 antibody revealed an intact endothelial cell monolayer in the uninjured control carotid artery (N = 3). As expected post balloon angioplasty, there is no endothelial cell monolayer of the injured left carotid arteries in the injury alone, injury + T-AuNP, and injury + S-AuNP treatment conditions. Green indicates autofluorescence of the elastic lamina. Magenta indicates endothelial cells. Images obtained using 25x magnification with exposure time of 400 msec. TUNEL staining for apoptosis revealed no evidence of endothelial cell apoptosis in the uninjured control carotid arteries, with a prominent endothelial cell monolayer (white arrowhead). Mild apoptosis was noted in the media of balloon-injured left carotid arteries, as expected. Black arrows indicate apoptotic cells. Representative images obtained using 40x magnification. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28242820), licensed under a CC-BY licence.

$KCl pulse decreased membrane, but increased nuclear, detection of NHE1 in b.End3 endothelial cells. (B) Representative immunoblots of NHE1, alpha-tubulin, PECAM, and lamin B in cytosol, membrane, and nuclear fractions of bEnd.3 cells harvested at 5 min after KCl or aCSF pulse. (C = cytosol, M = membrane, N = nuclear) Values represent the mean ratio of NHE1 detection +/- SEM (n = 11-12). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32469979), licensed under a CC-BY licence.$

TGF-beta1 released by MSCs contributes to induced myofibroblast differentiation and granulation tissue formationA-C2.5 x 105 of TGF-beta1 siRNA or control siRNA-transfected AT-MSCs were intradermally injected around each of CD18-/- murine wound. PBS mock injection served as negative control. Wound tissue was harvested at day 2, 5, and 7 post-wounding for quantification of human TGF-beta1 mRNA (A) at day 2 by qPCR, total TGF-beta1 (B), and active TGF-beta1 (C) protein at day 5 by ELISA. Data are expressed as mean +/- SEM, n = 3 wounds per group, **P < 0.01, by one-way ANOVA with Tukey's test.D-GExpression of alpha-SMA (D and E) and CD31 (F and G) at days 5 and 7 by immunostaining on tissue sections. The dashed lines indicate the border of the wound bed and epidermis or eschar. e, epidermis; es, eschar; wb, wound bed. Scale bars: 100 um. Quantification data are expressed as mean +/- SEM, n = 3 wounds per group, *P < 0.05, ***P < 0.001, by one-way ANOVA with Tukey's test.Source data are available online for this figure. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32080965), licensed under a CC-BY licence.

ATM is activated by spermidine in GM00637 cells.GM00637 cells with or without KU55933 pretreatment (a,b), and GM05849 cells (c) were exposed to 50 μM spermidine or CCCP, followed by immunofluorescence analyses of total and p-ATM on Ser-1981. The scale bar is 10 μm. Ratios of cells expressing p-ATM Ser-1981 to cells expressing total ATM were presented (d). 20–45 cells/condition from three experiments were collected. Values are mean ± SD, *p < 0.05 vs. control. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/srep24700), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Ddx19 interacts with MKL1 RPEL domain and Ipo beta .(a) Ddx19 recruitment from Hela cell lysate by GST-RPEL and GST-Ipo beta (left). WB, western blotting; (−), without Ran-Q69L; (+), with RanQ69L. Corresponding GST baits are stained with Ponceau to ensure the equal loading of the samples (below). Input sample corresponds to 5% of the Hela cell lysate used in the assay. (b) GST-RPEL was used as a bait for pull down of Ddx19 and Ipo beta from the Hela cell lysate in the presence of increasing amounts of LatB-actin (0.25–10 μM) (left). Arrow indicates the increasing amounts of actin in the corresponding Ponceau staining (below). Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/ncomms6978), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Ca

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ca

Applications:

, ,

Applications: , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

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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

Clone

Gene Symbol

Images Type

Isotype

Preservative

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Has Sample Size

Promotional Item

Species Glossary

LC3B Antibody - BSA Free

53BP1 Antibody - BSA Free

AIF-1/Iba1 Antibody

Collagen I Antibody

Carbonic Anhydrase IX/CA9 Antibody - BSA Free

GFP Antibody

Tyrosine Hydroxylase Antibody

Beclin 1 Antibody - BSA Free

TLR4 Antibody (76B357.1) - BSA Free

OGG1 Antibody - BSA Free

Laminin Antibody

CD31/PECAM-1 Antibody