Product Search for novus: Novus Biologicals

NB100-122

HIF-2 alpha/EPAS1 Antibody - BSA Free

(37 Reviews)

(781 Publications)

0.1 ml

NB100-122

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

Reactivity:

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

Species Glossary

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

Applications:

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

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

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

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

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-122SS

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NB100-479

HIF-1 alpha Antibody - BSA Free

(28 Reviews)

(497 Publications)

0.1 ml

NB100-479

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

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

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

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

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

Inhibition of Cdk1 and Cdk2 by CIP2A knockdown in cervical cancer SiHa cells caused G1 arrest. A, Western blot analysis of 16E6, p53 and CIP2A after HPV‐16E6 knockdown in cervical cancer SiHa cells. B, Protein expression of CIP2A, B‐Myb, Cdk1 and Cdk2 in SiHa cells after CIP2A knockdown. Data from a representative of 3 experiments are shown. C, Flow cytometric analysis of SiHa cells with CIP2A knockdown treated with PBS or bleomycin. A representative flow cytometry of 3 independent experiments is shown. D, Quantification of percentages G1 phase cells. Data from 3 experiments are summarized. *, P < .05 Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29893470), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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

Reactivity:

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

Species Glossary

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

Applications:

, , , , , , , ,

Applications: , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

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

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-479SS

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NB600-1384

LC3B Antibody - BSA Free

(19 Reviews)

(397 Publications)

0.1 ml

NB600-1384

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

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

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

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

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

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

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

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

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

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

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

LC3B staining in glioblastoma multiform tissue.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

Reactivity:

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

Species Glossary

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

Applications:

, , , , , , , , , ,

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

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

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

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB600-1384SS

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AF2018

SOX2 Antibody [Unconjugated]

(6 Reviews)

(350 Publications)

100 μg

AF2018

Western blot shows lysates of D3 mouse embryonic stem cell line, NTera-2 human testicular embryonic carcinoma cell line, F9 mouse teratocarcinoma stem cells, and rat cortical stem cells. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse/Rat SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for SOX2 at approximately 36 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

BG01V human embryonic stem cells were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. SOX2/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human/Mouse/Rat SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) or control antibody (AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (MAG999) and DNA was purified using chelating resin solution. Thenanogpromoter was detected by standard PCR.

SOX2 was detected in immersion fixed undifferentiated mouse cortical stem cells using Goat Anti-Human/Mouse/Rat SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; (NL001) and counterstained with DAPI (blue). Nestin was also detected in stem cells using Mouse Anti-Mouse/Rat Nestin Monoclonal Antibody (MAB2736) and co-stained using the NorthernLights™ 493-conjugated Anti-Mouse IgG Secondary Antibody (green; NL009). Specific staining of SOX2 was localized to nuclei. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.

SOX2 was detected in immersion fixed undifferentiated rat cortical stem cells using Goat Anti-Human/Mouse/Rat SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Nestin was also detected using Mouse Anti-Mouse/Rat Nestin Monoclonal Antibody (MAB2736) and stained using the NorthernLights™ 493-conjugated Anti-Mouse IgG Secondary Antibody (green; NL009). Specific staining of SOX2 was localized to nuclei. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.

Simple Western lane view shows lysates of BG01V human embryonic stem cells and NTera-2 human testicular embryonic carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for SOX2 at approximately 53 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse/Rat SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

SOX2 was detected in immersion fixed ADLF1 (top panel) and FAB2 (bottom panel) induced pluripotent stem cell lines using Goat Anti-Human/Mouse/Rat SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.

SOX2 was detected in immersion fixed paraffin-embedded sections of glioblastoma using Goat Anti-Human/Mouse/Rat SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei in cancer cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

SOX2 was detected in immersion fixed NTera‑2 human testicular embryonic carcinoma cells (Positive) & absent in HepG2 human hepatocellular carcinoma cells (Negative) using Goat Anti-Human/Mouse/Rat SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to Cell surface and nuclear. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Knockdown OGT inhibits proliferation and tumorsphere formation of hepatoma cell through reducing eIF4E expression. A, Huh7 and PLC/PRF/5 cells were infected with control shRNA, OGT shRNA2 alone, or with wild‐type eIF4E lentivirus. The cell lysates were harvested for western blotting analysis using indicated antibodies. beta ‐actin expression was served as a loading control. B, Cell proliferation of Huh7 and PLC/PRF/5 cells infected with lentiviruses as in panel (A) were measured with CCK8 assay. (C‐H) Huh7 and PLC/PRF/5 cells infected with lentiviruses as in panel (A) were seeded into 96‐well plates. After 12 d, tumorsphere were counted and quantified. Representative images of sphere (scale bars, 100 μm) were shown (C, F). The diameter of sphere (D, G) and number of sphere (E, H) were count. Data represent mean ± SD of at least three independent experiments. The two‐tailed Student's t tests were used. **P < 0.01. I, Huh7 cells expressing either OGT shRNA2 alone or with wild‐type eIF4E lentivirus were incubated with PE‐labelled anti‐AC133 antibody. The percentages of CD133+ cells in graphs were analysed by flow cytometry. Black line, control IgG staining; red line, CD133 staining. J, Cell lysates were examined by western blotting with indicated antibodies. The right panel showcases relative protein amounts of different groups. Error bars represent ±SD of triplicate experiments. **P < 0.01; n.s, no significance. K, Huh7 cells were collected and subjected to immunoprecipitation with antibody against eIF4E or normal mouse IgG. Total RNAs were purified from immunocomplexes and subjected to RT‐PCR to measure Sox2, OCT4, and KLF4 mRNAs associated with eIF4E Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30677218), licensed under a CC-BY license. Not internally tested by R&D Systems.

TP63 and SOX2 staining in cervical biopsies.Immunohistochemical staining of cervical biopsies. Bars = 50µm.(A) normal cervix no primary control; (B) normal cervix stained with anti-SOX2; (C) Squamous cell cervical carcinoma no primary control; (D) representative TP63, and (E) representative SOX2, staining of tumour cells. For both TP63 and SOX2 staining was seen in the nucleus of positive cells (examples indicated by solid arrows); negative cells were a minority of tumour cells (examples indicated by unfilled arrows). (F) Image analysis results of % nuclear +ve tumour cells in biopsies. Parallel sections from 11 cases were stained with SOX2 and TP63. Tumour cells were evaluated for their nuclear expression of the transcription factors. There was no significant difference between the data for SOX2 and TP63 (Wilcoxon signed rank test). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25531390), licensed under a CC-BY license. Not internally tested by R&D Systems.

Direct effects of glucocorticoids in ependymal glia. (A–I) Zebrafish spinal cord transverse sections. (A–C) Immunostaining of wild-types for Nr3c1 (green) and Gfap (red), plus DAPI staining (blue) with Dex treatment at 24 h post sham injury (A), 24 h (B) and 48 h post SCI (C). Nuclear Nr3c1 (active) localization in Gfap-positive cells around the central canal is stimulated with Dex treatment. (D–F) Immunostaining for EGFP (pan-cytoplasmic) in the SR4G transgenic reporter line (green) and filamentous Gfap (red) at 24 h post sham injury (D) and SCI with no treatment (E) or 24 h post SCI +Dex (F). Nr3c1 signaling is constitutively active in ependymal glia in uninjured controls (D), diminished after SCI (E), and was maintained after SCI + Dex. (G–I) Immunostaining for Sox2 (green), PCNA (red) and Gfap (blue) from 48 h post sham injury (G), 48 h post SCI followed by no treatment (H), or 48 h post SCI +Dex (I). Sox2 immunoreactivity is reduced in ependymal glia and neural precursor cells after Dex treatment. Representative images from 10 spinal cords per time point in each condition. Scale bars, 20 μm (A is the same as B,C; D is the same as E,F; G is the same as H,I). ∗denotes central canal. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31069223), licensed under a CC-BY license. Not internally tested by R&D Systems.

Epigenetic perturbation of enhancer clustering and genome-widebinding.(A) The fluctuation range (x) and amplitude (y) were obtainedby fitting the pair-correlation function of the indicated dataset with thefluctuation model. Figure 2 and Figure 2—figure supplement 1,Equations10–13. Supplementary file 1. Data from the same conditionwere grouped in separate ellipses. (B) Sox2 ChIP-exo peakdensity distribution in the wild-type and TSA treated (red dotted) cellsacross chromosome 1, 2, 3. In the upper panels, each chromosome was dividedto 500 bins. The color map correlates with the number of peaks in each bin.Top 7000 binding sites were considered in each condition. (C)Cumulative density histogram of the distances to transcription start sites(TSS's) of Sox2 ChIP-exo peaks in WT, Sox2 ChIP-exo peaks in the TSA treatedcells (red dotted), and random genomic positions (gray).DOI://dx.doi.org/10.7554/eLife.04236.013 Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25537195), licensed under a CC-BY license. Not internally tested by R&D Systems.

Generation of KCNA5 knockout model. (A) Illustration of polycistronic self-replicating RNA used for reprogramming (Yoshioka et al., 2013). Reading frames for OCT4, KLF4, SOX2, GLIS1, and puromycin resistance are separated by 2A peptide-encoding or IRES sequences. (B) Phase contrast morphology (5 x) and alkaline phosphatase staining of the F1 wild-type hiPSC line generated using the RNA vector shown in (A). (C) Representative F1 karyogram indicating a normal male genotype. (D) The RNA vector is undetectable in established F1 hiPSCs following puromycin withdrawal, whereas freshly transfected parental fibroblasts display robust transgene expression (RT-qPCR data, n = 2). (E) Bisulfite sequencing of OCT4 and NANOG promoter regions of the indicated samples showing an hESC-like hypomethylated state in F1 cells. (F) RT-qPCR analysis of F1 hiPSCs in comparison to the original fibroblasts and a hESC control (n = 2). (G) Immunostains showing robust pluripotency factor expression in F1 hiPSCs at the protein level. (H) Illustration of the Kv1.5 channel. The voltage sensor in transmembrane helix S4 is indicated by symbols. The ion pore region is in-between segments S5 and S6. (I)KCNA5 knockout strategy based on simultaneously transfecting 4 CRISPR/Cas9 nickase vectors targeting the 5' end of the 1-exon gene body. A mutant clone generated this way displayed homozygous deletions causing a frame shift and premature stop as illustrated at the bottom. (J) PCR spanning the mutation site validates the sequencing-verified frame shift-causing deletion in KCNA5fs/fs cells at the DNA and RNA levels. Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fphys.2017.00469/full), licensed under a CC-BY license. Not internally tested by R&D Systems.

Construction and expression of srRNA.A: Scheme of Self-Replicating srRNA constructs. B: Expression of reprogramming factors in OKS-iM 4F-srRNA, OKS-iG 4F-srRNA, OKS-iGM 5F-srRNA, and OKS-iGML 6F-srRNA on day 2 (left) and day 10 (right). BJ cells were co-transfected with srRNAs plus B18R mRNA (1:1 srRNA:B18R mRNA), selected with puromycin and cultured in Advanced DMEM/10% FBS for 10 days. C: qRT-PCR analysis of LIN28A and NANOG from 4F-srRNA and 5F-srRNA transfected BJ cells normalized to GAPDH. Cells were selected with puromycin after the transfection, and cultured in Advanced DMEM/10% FBS (Ad) or ES medium. Cells were collected on day 2, 4, 6, 8 and 10 for 5F-srRNA, and day 10 for 4F-srRNA (OKS-iM and OKS-iG). D: Immunoblot analysis of LIN28A and NANOG in 5F-srRNA transfected BJ cells on day 6 and 10. iPSC: iPSC clone generated with 5F-srRNA from BJ cells. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28750082), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , , ,

Applications: , , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Biotin	BAF2018
NL493	NL20181G

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	NL20181G
Carrier Free	AF2018-SP

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NB100-142

Mre11 Antibody

(4 Reviews)

(234 Publications)

0.05 ml

NB100-142

<b>Genetic Strategies Validation. </b>Knockdown Validated: Mre11 Antibody [NB100-142] - Mre11 and NBS1 contribute to OriP replication and EBV episome stability. D98/HR1 cells were transfected with siRNA for Mre11 or NBS1, or Luciferase control and assayed by immunoblot (IB) with antibodies specific for Mre11 (left pane) or NBS1 (right panel), or PCNA (lower panel) as a loading control. Image collected and cropped by CiteAb from the following publication (//journals.plos.org/plosone/article?id=10.1371/journal.pone.0001257) licensed under a CC-BY license.

Immunohistochemistry: Mre11 Antibody [NB100-142] - Immunohistochemical staining of MRE11 in a human epidermis cross section.

Western Blot: Mre11 Antibody [NB100-142] - Western blot analysis of Mre11 on 50 ug of HeLa and MEF lysates, displaying bands at the molecular weight of 81 kDa.

Immunocytochemistry/Immunofluorescence: Mre11 Antibody [NB100-142] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-Mre11 [NB100-142] at a 1:500 dilution overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Chromatin Immunoprecipitation: Mre11 Antibody [NB100-142] - Aggregated END-seq signal and MRE11 ChIP-seq RPM in WT (left) and Atm -/- (right). To fairly compare ChIP-seq signal between WT and Atm -/-, MRE11 scale is proportional to spike-in normalized END-seq RPM for each genotype. Individual hotspot examples (chr12:34,592,264-34,598,265) are shown below. Note that decreased MRE11 coverage is observed within NDR of Atm -/-. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32051414/) licensed under a CC-BY license.

Knockdown Validated: Mre11 Antibody [NB100-142] - TIG-1 human primary fibroblasts, whole cell lysate (30 ug). Image from verified customer review.

ATM coordinates multiple levels of resection machinery. d Aggregated END-seq signal and MRE11 ChIP-seq RPM in WT (left) and Atm-/- (right). To fairly compare ChIP-seq signal between WT and Atm-/-, MRE11 scale is proportional to spike-in normalized END-seq RPM for each genotype. Individual hotspot examples (chr12:34,592,264-34,598,265) are shown below. Note that decreased MRE11 coverage is observed within NDR of Atm-/-. e Aggregate plot overlapping WT END-seq and Atm-/- MRE11 ChIP-seq, normalized to the same height. MRE11 shows prominent localization to the WT END-seq read-less gap. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32051414), licensed under a CC-BY licence.

Elevated levels of various proteins in MEC and in dentate gyrus of MPS IIIA brain.Staining was performed with antibodies against the proteins shown in a 7 months-old MPS IIIA mouse brain. The first 3 rows are for the MEC region and the bottom row for dentate gyrus. Age-matched C57BL6 mice, used as controls, showed no staining in the MEC region (not shown). The dentate gyrus showed AT270 inclusions in MPS IIIA (−/−) mouse brain (arrows) but not in the C57BL/6 (control) brain (bottom row). 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.

$SUMO2-PCNA enriches CAF1 and FACT within the replication complex. a Western blot analysis of the indicated proteins in the CB fractions (left) and the FLAG-PCNA complexes purified from CB fractions (right) prepared from cells expressing the indicated FLAG-tagged proteins. Ub-KR indicates the FLAG-ubiquitin-PCNA (KR) fusion protein. b Western blot analysis of the indicated proteins in the CB input (left panels) used for the in vitro pull-down experiments of purified recombinant strep-tagged PCNA, SUMO2-conjugated PCNA, and GFP anchored to Strep-Tactin resins (right panels). SUMO2-modified PCNA was generated by the in vitro SUMOylation reaction using strep-PCNA and His-SUMO2 shown in Supplementary Fig. 6a, b. c Western blot analysis of the input levels of recombinant strep-PCNA and strep-SUMO2-PCNA fusion (S2-PCNA) purified from E. coli (left panel) and of strep-PCNA and strep-S2-PCNA pulled down by WT or SIM mutant FLAG-CAF1A proteins purified and anchored on FLAG M2 agarose beads from HEK293T cells expressing the corresponding constructs (right panels). d Western blot analysis of strep-S2-PCNA (upper 2 panels) and strep-PCNA (lower 2 panels) pulled down by FLAG-CAF1A purified from E. coli and anchored on FLAG M2 agarose beads. e Western blot analysis of the indicated proteins in the CB input prepared from HEK293T cells expressing FLAG-SSRP1 WT or SIM mutant (left panels) and of FLAG-SSRP1 and SPT16 pulled down from the CB fractions in vitro using purified recombinant strep-tagged PCNA, S2-PCNA, and GFP anchored to Strep-Tactin resins (right panels) Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30006506), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Restoration of nuclear PTEN-mediated DNA damage signalling by restraining CK2 impairs tumour but not normal neural stem cell survival.(a) Immunoblot analysis displaying the impact of TBB treatment (25 μM/24 h) on CK2 beta , PTEN, p-PTEN (S380/T382/T383), p-AKT (S473) and p-PRAS40 (T246) in GXP GBM3 & 4, with beta -actin used as loading control. (b) IF of PTEN with DAPI nuclear staining in dimethylsulfoxide (DMSO)- or TBB (25 μM/24 h)-treated cells in the absence of DOX or after 5 h of exposure to DOX (0.5 μM). DOX was added 5 h before the end point of DMSO or TBB treatment. Representative images are shown. Scale bars, 20 μm. (c) Co-IF of gamma H2AX and 53BP1 foci formation with DAPI nuclear staining in DMSO- or TBB (25 μM/24 h)-treated GXP NS or GBM2 with DOX−, or 1 h or 5 h of DOX (0.5 μM) exposure followed by 5 or 24 h of recovery time. Scale bars, 10 μm. A graph quantifying the percent of gamma H2AX-positive GXP NS or GBM2 cells (y axis) with DMSO or TBB treatment at indicated time points of DOX treatment (x axis) is summarized in the graph in (d). Error bars represent mean±s.d. Representative 100 cells were randomly selected for quantification. Cells with ≥5 foci were considered as gamma H2AX positive. (e) Co-IF of FANCD2 and gamma H2AX with DAPI nuclear staining in TBB (25 μM/24 h)-treated GXP GBM2 exposed to DOX (0.5 μM) 5 h before fixation. Scale bars, 10 μm. The percent of FANCD2/ gamma H2AX-positive foci at DSB sites (y axis) in TBB− or TBB+ (x axis) in GXP GBM2 is summarized in the graph shown in (f). Error bars represent mean±s.e.m. See also Supplementary Fig. 4. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28094268), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

GRPs enhance axonal growth in DRGs in vitro. A–C, Representative montage images are shown for control dissociated DRG culture (A), dissociated DRGs cocultured with rat GRPs (B), and dissociated DRGs cultures exposed to conditioned medium from alkaline phosphatase-expressing rat GRPs (C). beta III-tubulin (red) immunofluorescence highlights the neurons; GRPs are visualized by immunostaining for alkaline phosphatase (green). D shows quantification for the average length of the longest axon per neuron ± SEM (n ≥ 30 neurons in three separate experiments; **p ≤ 0.01 by Student’s t test). Scale bar, 250 μm. Image collected and cropped by CiteAb from the following open publication (//www.eneuro.org/lookup/doi/10.1523/ENEURO.0171-16.2017), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Ch, Ha, Bt, Bv, Ca, Pm, Eq, Fe, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, Ch, Ha, Bt, Bv, Ca, Pm, Eq, Fe, Pm

Applications:

, , , , , , , , , , ,

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

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Validated by:

Validated by:

Available Formulations

Formulation	Catalog #	Availability	Price
Purified	NBP3-25349-0.025mg
Purified	NBP3-25349-0.05mg

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NBP1-06712

DYKDDDDK Epitope Tag Antibody (L5)

(10 Reviews)

(135 Publications)

0.5 ml

NBP1-06712

Western Blot: DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] - Analysis of DYKDDDDK tagged AIFm2 protein (A), DYKDDDDK tagged IREB2 protein (B), and vector control (C) using DYKDDDDK epitope tag antibody (L5) [NBP1-06712] at 2 ug/ml.

Immunocytochemistry/Immunofluorescence: DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] - Analysis of HeLa cells transfected with a FLAG encoded construct (top) or untransfected (bottom), using DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712]. Image from verified customer review.

Immunohistochemistry: DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] - Third instar larva of the genotype [hs-Flp; nrv2-Gal4/MCFO-2]. flp expression of the multicolor FlpOut construct was induced by 1 h 37C heat shock during first instar stage. Larvae were stained for HA (green), V5 (red), and FLAG (blue). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32901033/) licensed under a CC-BY license.

Western Blot: DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] - Detection of DDK in SUSD3 overexpression lysate using [NBP1-06712] followed by goat anti-rat IgG HRP conjugated secondary antibody (cat.# NB7115). Image from verified customer review.

Immunocytochemistry/Immunofluorescence: Rat Monoclonal DYKDDDDK Epitope Tag Antibody (L5) [NBP1-06712] - Analysis using DyLight 488 conjugated antibody (NBP1-06712G). Normal mouse prostate cells overexpressed with Flag-tagged construct (Green: Flag). Image from verified customer review.

$SHP inhibits AhR transactivation of Pemt in response to feeding or FGF19 treatment. a, b Hepa1c1c7 cells were grown in low-glucose and serum-free media for 12 h, and transferred for 15 min to complete medium or treated for 15 min with insulin, FGF19, or CDCA, and then cells were harvested. Cells were pre-treated with a PKB inhibitor, PKB124005, or an ERK inhibitor, PD98059, for 30 min prior to insulin treatment as indicated. Levels of AhR and SHP in the cytoplasmic and nuclear fractions were determined by IB. Consistent results were observed from two independent studies. Full size immunoblots are in Supplementary Figure 12a. c Hepa1c1c7 cells were treated with insulin for 15 min or 45 min, and pre-mRNA levels of Pemt and Cyp1a1 were measured (n = 6). d Mice were fasted for 12 h (Fs) or fasted and refed for 4 h (Fd) or fasted 4 h and treated with FGF19 for 2 h, liver nuclear extracts were prepared, and the interaction of SHP and AhR was examined by CoIP. e, f Mice were fasted overnight (Fs) or fasted and refed for 6 h (Fd). e Cellular localization of AhR and SHP in mouse liver sections was examined by IF. Scale bar: 100 µM. f Liver chromatin was immunoprecipitated with AhR antibody, then eluted, and re-precipitated with ARNT or SHP antibody to examine the occupancy of SHP at AhR-bound chromatin at the Pemt promoter (n = 5 mice). g Hepa1c1c7 cells were transfected with the indicated plasmids, and then treated with FGF19 for 2 h and luciferase activity was determined (n = 5). h, i Mice (n = 5 mice per group) were fasted for 12 h and then refed for the indicated times, and the occupancy of AhR, ARNT, and SHP at the Pemt promoter (h) or Pemt pre-mRNA levels (i) were determined by liver ChIP assay or by qRT-PCR, respectively. Means ± SD are shown, and statistical significance was measured using the c, g, i one- or f, h two-way ANOVA with the FDR post-test. *P < 0.05, **P < 0.01, NS, statistically not significant Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29416063), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Reactivity:

ET

Species Glossary

Reactivity: ET

Applications:

, , ,

Applications: , , ,

Host:

Rat Monoclonal

Host: Rat Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP1-06712AF350
Alexa Fluor 405	NBP1-06712AF405
Alexa Fluor 488	NBP1-06712AF488
Alexa Fluor 532	NBP1-06712AF532
Alexa Fluor 594	NBP1-06712AF594
Alexa Fluor 647	NBP1-06712AF647
Alexa Fluor 700	NBP1-06712AF700
Alexa Fluor 750	NBP1-06712AF750
Biotin	NBP1-06712B-0.1ml
DyLight 350	NBP1-06712UV
DyLight 405	NBP1-06712V
DyLight 488	NBP1-06712G-0.1ml
DyLight 550	NBP1-06712R-0.1ml
DyLight 594	NBP1-06712DL594
DyLight 650	NBP1-06712C-0.1ml
DyLight 680	NBP1-06712FR
DyLight 755	NBP1-06712IR
FITC	NBP1-06712F-0.1ml
HRP	NBP1-06712H-0.1ml
Janelia Fluor 549	NBP1-06712JF549
Janelia Fluor 646	NBP1-06712JF646

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NB100-134

HIF-1 alpha Antibody - BSA Free

(14 Reviews)

(220 Publications)

0.1 ml

NB100-134

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody - BSA Free [NB100-134] - Analysis of HIF-1 alpha in zebrafish retina tissue using NB100-134 HIF-1 alpha antibody. Mouse tissue was used as a control. Image from verified customer review.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-134] - Analysis of HIF-1 alpha in overexpression and endogenous HIF-1 alpha & HIF-2 alpha using anti-HIF-1 alpha antibody NB100-134. The data showed that HIF-1 alpha antibody did not react to HIF-2 alpha overexpression. Image from verified customer review.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-134] - Analysis of HIF-1 alpha using NB100-134, on normoxic and hypoxic nuclear rat cell lysates.

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody - BSA Free [NB100-134] - Representative images of macrophages and activated macrophages, in the absence or presence of 10 uM hemin for 72 h. HIF-1 alpha levels were detected by immunofluorescence using a confocal imaging system (Opera Phenix, Perkin Elmer). Images were acquired with 20x water objective. Staining intensity levels in the nucleus and cytosolic region were obtained using Harmony software (Perkin Elmer). Nucleus and cytosol were identified through Hoechst (at 405 nm) and CellMask (at 655 nm) staining, respectively. Scale is shown as 100 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33187129/) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody - BSA Free [NB100-134] - Analysis of HIF-1 alpha in Normoxic (N) and Hypoxic (H) ARPE-19 cells using anti-HIF-1 alpha antibody NB100-134. Image from verified customer review.

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody - BSA Free [NB100-134] - Hif1alpha expression in 9.5dpc primordial germ cells. Whole-mount staining of Hif1alpha and GFP in Oct4-GFP embryo (lateral view). Scale bar: 100um. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0154309) licensed under a CC-BY license.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody - BSA Free [NB100-134] - Staining in the canine CL on days 10 to 70 after ovulation. PC = positive control (human placenta). NC = negative control. Image from verified customer review.

Immunohistochemistry: HIF-1 alpha Antibody - BSA Free [NB100-134] - Staining of human kidney, renal tubular epithelium in cortex using NB100-134.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody - BSA Free [NB100-134] - Analysis of FFPE tissue section of human endometrium carcinoma AN3CA cell line based xenograft using rabbit polyclonal HIF-1 alpha antibody NB100-134 at 1:300 dilution. The signal was developed using HRP-labelled secondary antibody and DAB reagent, and the section was further counterstained using hematoxylin. The tested section depicted mainly a diffused cytoplasmic staining but there were some cells which showed nuclear signal also (representing hypoxic cells).

Simple Western: HIF-1 alpha Antibody - BSA Free [NB100-134] - Image shows a specific band for HIF-1 alpha in 0.2 mg/mL of Hypoxic HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Immunoprecipitation: HIF-1 alpha Antibody - BSA Free [NB100-134] - PC-3 cell lysates run with cobalt chloride treatments. Immunoprecipitation was performed with protein A/G agarose beads. This data was provided courtesy of Kelie Reece, Figg lab, NCI.

Dual RNAscope ISH-IHC: HIF-1 alpha Antibody - BSA Free [NB100-134] - Formalin-fixed paraffin-embedded tissue sections of human kidney were probed for HIF-1 alpha mRNA (ACD RNAScope Probe, catalog #605228; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using Rabbit Polyclonal (Novus Biologicals catalog # NB100-134) at 1:500 dilution with one-hour incubation at room temperature followed by incubation with anti-rabbit IgG VisUCyte HRP Polymer Antibody (Catalog # VC003) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm of tubules.

Simple Western: HIF-1 alpha Antibody - BSA Free [NB100-134] - Simple Western lane view shows lysates of MCF +/- DMOG, BioSpherix MSCs in hypoxic conditions, and BioSpherix MSCs in normoxic conditions loaded at 0.5 mg/ml. A band was detected for HIF-1 alpha at approximately 116 kDa (as indicated) using NB100-134 (1:100 dilution) followed by Anti-Rabbit Secondary Antibody (042-206, ProteinSimple). This experiment was conducted under standard assay conditions, and using the 12-230 kDa separation module (SW-W004). Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody. Image from an internal validation.

P53 was increased response to IR in the fetal brain.Even though no significant change in protein level, total p53 and phosphorylation of P53 at ser15 were increased compared with HIF-1 alpha in fetal brains response to IR. Numbers in the pictures show the average of three independent experiments (n = 3 from 3 individual pregnant mice).

IGFBP5 inhibits HIF1alpha expression through the MAPK-ERK signaling pathway. C). Overexpression of IGFBP5 inhibited HIF1alpha expression visualized by western blots of A375 cells. The arrow points to the band of HIF1alpha. D. Assessment of the reduced gene expression levels of VEGFA and MMP9, downstream genes regulated by HIF1alpha, in A375 IGFBP5 OE compared to control cells by qRT-PCR analysis. Data were shown for the mean +/- SD form three independent experiments. P values based on two-side Student t-test comparing A375 IGFBP5 OE tumor cells and vector control cells. *, P 25 pictures/group from three separate experiments. Scale bar = 40 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31083380), licensed under a CC-BY licence.

ER-INP binds to PHD2, inhibits hydroxylation of HIF-1alpha and increases HIF-1alpha accumulation. (B) Western blot analysis to measure the effect of ER-INP on HIF-1alpha and its hydroxylation level in transfected HEK293 and RAW264.7 cells pre-treated with MG132 (upper panel) and the protein ratio to beta-actin loading control by ImageJ densitometry analysis (lower panel; n = 3). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31413262), licensed under a CC-BY licence.

HIF-1alpha levels are increased by androgen stimulus and Tip60 overexpression in LNCaP cells. (A) Nuclear and (B) cytosolic HIF-1alpha levels and (C) images in LNCaP cells and in LNCaP cells overexpressing Tip60, in the absence or presence of androgen (10 nM R1881, 72 h). HIF-1alpha levels were detected by immunofluorescence using confocal imaging system. Images were acquired with 20x objective. Staining intensity levels in the nucleus and cytosolic region were obtained using Harmony software. Nucleus and cytosol were identified through Hoechst and CellMask staining, respectively. Scale is shown as 100 um. White dotted frames indicate the section of the image that was enlarged. Values are expressed as mean +/- SEM, from three independent culture preparations, each treatment performed in quadruplicate. Two-way ANOVA, Bonferroni post-test and p values comparisons are specified in the figures (* p < 0.05). HIF-1alpha, hypoxia-inducible factor-1alpha; OE, overexpressing; R1881, synthetic androgen. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31671779), licensed under a CC-BY licence.

The increase of cytosolic HIF-1alpha levels of activated macrophages is inhibited by hemin. (C) Representative images of macrophages and activated macrophages, in the absence or presence of 10 uM hemin for 72 h. HIF-1alpha levels were detected by immunofluorescence using a confocal imaging system (Opera Phenix, Perkin Elmer). Images were acquired with 20x water objective. Staining intensity levels in the nucleus and cytosolic region were obtained using Harmony software (Perkin Elmer). Nucleus and cytosol were identified through Hoechst (at 405 nm) and CellMask (at 655 nm) staining, respectively. Scale is shown as 100 um. White dotted frames indicate the section of the image that was enlarged. Values with different superscript letters (a and b) indicate significant differences (p < 0.05) between groups. For all bars with the same letter, the difference between the means is not statistically significant. HIF-1alpha, hypoxia-inducible factor-1 alpha. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33187129), licensed under a CC-BY licence.

LOXL2 abundance in EC-derived exosomes is increased in hypoxia. (A) Immunoblots of endothelial cells for HIF-1alpha (control for hypoxia, top panel), LOXL2 (middle panel) and beta-actin (loading control, lower panel). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26612622), licensed under a CC-BY licence.

Hypoxic setup for larval zebrafish. (D) Western blot of Hif1alpha protein expression showing that hypoxia enhanced Mbp protein translation. *P < 0.05; **P < 0.01; ***P < 0.001. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30337858), licensed under a CC-BY licence.

Enhanced expression of hypoxia-induced survival factors and angiogenic growth factors in hASC L-spheroids. (C) Western blot analysis and quantification of HIF1-alpha in hASCs cultured as spheroids, L-spheroids and monolayers (*p < 0.01, compared to the L-spheroid group). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26065900), licensed under a CC-BY licence.

E2 enhances neutrophil apoptosis through suppressing the expression of hif1alpha and c-myb under physiological conditions. C) Staining of hif1alpha with antibody with or without E2 treatment. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35842445), licensed under a CC-BY licence.

HIF1alpha and AhR mediate butyrate induction of IL-22 in CD4+ T cells.. b-f CD4+ T cells were activated with anti-CD3/CD28 mAbs +/- butyrate (0.5 mM) under Th1 conditions (n = 3/group). Hif1a (b) and Ahr (c) were analyzed by qRT-PCR. HIF1alpha (d) and AhR (e) protein was analyzed by western blot on day 2. HIF1alpha activity was measured using HIF1alpha Transcription Factor Assay Kit (f). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32901017), licensed under a CC-BY licence.

$Alteration of PARKIN leads to an impairment of mitophagy upon CCCP treatment.a 2 N and DS were treated with CCCP (20 μM) for 6 h and mitochondrion/cytosol fractionation was performed to assay the accumulation of LC3-II and p62 within the mitochondrial fraction. VDAC1 and ACTIN were used as a loading control for mitochondrial and cytosolic fractions, respectively. b, c Quantitative data of LC3-II and p62 in the mitochondrial fraction were normalized with VDAC1 (n = 3). d The cells were treated with CCCP (20 μM) for 6 h and the whole-cell extracts from 2 N and DS were analyzed by western blot for PARKIN and PINK1. e Quantification of PARKIN normalized with ACTIN (n ≥ 3). d Quantification of PINK1 after 6 h of CCCP, each quantitative data was normalized with ACTIN (n ≥ 3). g Representative confocal images of cells stained with Mitotracker green (mitochondrial marker) and Lysotracker red (lysosomal marker) upon CCCP treatment (20 μM for 6 h). White arrow heads point to the co-localization within puncta from the boxed region at high magnification shown on the right. Scale bar 10 μm. h The graph shows the percentage of Mitotracker green /Lysotracker red colocalization calculated by JACoP plugin of ImageJ. Minimum 50 cells per condition were counted from three independent experiments. i 2 N and DS cells were treated with CCCP (20 μM) for 6 h and were assessed for SQSTM1 (p62), PARKIN and PINK1 mRNA by quantitative real-time PCR. mRNA levels were normalized to ACTIN mRNA, used as internal control. Data display the fold-changes of SQSTM1, PARKIN and PINK1 mRNAs relative to control cells (n = 3). Statistical analysis was performed using Student’s T-test or one-way ANOVA with Tukey’s multiple comparisons test. (*p < 0.05; **p < 0.01; ***p < 0.001; n.s. no significant difference). Immunoblots reported are from one representative experiment Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31332166), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

WNT11 is induced by hypoxia or hypoxic mimetics in different cell types.(A) Increased Wnt11 mRNA in EMSC adipocytes (Day 12) after hypoxia-mimetic treatments. EMSC adipocytes were treated with CoCl2 (0.1 mM), DFO (0.1 mM) or DMOG (0.1 mM) for 24 hrs. Values were normalized to Tbp mRNA and are expressed relative to control (n = 3). (B,C) Increased Wnt11 mRNA by hypoxia in EMSC preadipocytes and adipocytes (Day 0–12 after differentiation) (B), and C2C12 myoblast and myocyte (Day 0 and 8 after differentiation) (C). Wnt11 mRNA was assessed by quantitative PCR in cells exposed to air (21% O2) or hypoxia (1% O2) for 24 hrs. (n = 4). Values were normalized to Tbp mRNA and are expressed relative to 21% O2 samples (left panel). (D) Immunoblot analyses of HeLa cells under normal air or hypoxia for 24 hrs. (E,F) Induction of Wnt11 by increasing concentrations of DMOG in MDA-MB-231 cells (E) and 4T1 cells (F). (G) EMSCs treated with 0.1 mM DMOG for the indicated times. Wnt11 and Vegf mRNA expression was measured by qPCR and normalized to Tbp mRNA (n = 4). (H) WNT11 protein levels after DMOG treatment normalized to alpha -Tubulin (upper panel; n = 4). Representative immunoblots of EMSCs treated with 0.1 mM DMOG for the indicated times (Lower panel). (I) Protein expression in MDA-MB-231 cells treated with 0.1 mM DMOG. (J) Induction of Wnt11 promoter activity by hypoxia or hypoxia mimetics. pGL3-Wnt11 promoter plasmid was transfected into C2C12 cells. Cells were incubated with DMOG (left panel, n = 4) or under 21% O2 or 1% O2 (right panel, n = 8) for 24 hrs. For panels (A–C,G,H,J), values are mean ± s.e.m. *p < 0.05, **p < 0.01. For panels of immunoblotting, laminin, alpha -tubulin, and ERK were used as loading controls, WNT11 normalized to alpha -Tubulin was shown. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/srep21520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Metformin synergized with sorafenib to suppress angiogenesis, proliferation, and EMT but promote apoptosis in vivo. Representative images of tumor sections were stained with antibody against CD31, Ki67, and TUNEL (magnification ×200). a Metformin in combination with sorafenib synergistically minimized both Ki67 and CD31 expression but promoted TUNEL expression. b Combined therapy inhibited EMT at protein level which lysated from the tumor tissue, and pAMPK expression was elevated both in metformin group and combined treatment groups. c Combined treatment of metformin and sorafenib suppressed HIF-2 alpha protein expression but upregulated TIP30 protein expression evaluated by Western blot Image collected and cropped by CiteAb from the following open publication (//www.jhoonline.org/content/9/1/20), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

EBNA1 expression is associated with upregulation of MTH1 and oxidative damage repair pathways. The expression of MTH1, OGG1, and MUTYH was investigated by western blots and qPCR in BJAB-tTAE1 cells upon addition and withdrawal of doxycycline. a Representative western blots illustrating the correlation between the reversible down- and upregulation of MTH1 and EBNA1 in BJAB-tTAE1 cells upon addition and withdrawal of doxycycline. GAPDH was used as loading control. b Densitometry quantification of MTH1 and EBNA1 expression in BJAB-tTAE1 cells. The mean intensity of the MTH1 and EBNA1 specific bands relative to GAPDH in three independent experiments is shown in the figure. c Regression analysis of the relationship between expression levels of MTH1 and EBNA1. The data from three independent experiments were used for the plot. d Representative western blots illustrating the expression of MTH1, MUTYH, and OGG1 in BJAB-tTAE1 cells cultured for two weeks in the presence or absence of doxycycline. e Fold change is the ratio between the intensity of the specific band in cells cultured without or with doxycycline. The mean ± SD of four independent experiments is shown. f qPCR analysis of the levels of MTH1, OGG1, and MUTYH transcripts in BJAB-tTAE1 cells cultured for 2 weeks in the presence or absence of doxycycline. The mean ± SE of the fold change in six independent experiments each performed in triplicate is shown 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.

Impacts of PRDX3 protein on autophagy flux(A–D) Representative immunoblot (A, C) and quantification (B, D) showing the levels of LC3-II in BPH-1 cells treated with random (MOCK) or PRDX3-specific siRNA (PRDX3) (A, B) or RWPE-1 cells transiently expressing different amount of PRDX3 (C, D) in the absence (Ctrl) or presence of bafilomycin A1 (BAF). Data are mean and standard deviation of three repeats and differences are tested with Student's T-test. *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001. (E–G) Representative immunoblot (E) and quantification (F, G) showing the levels of Beclin 1 (F) and PI3KCIII (G) in BPH-1 cells treated with random (MOCK) or PRDX3-specific siRNA (PRDX3). Ns, not significant; *P ≤ 0.05. Image collected and cropped by CiteAb from the following open publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.17927), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Autophagy level was promoted by downregulation of the Hh signaling pathway in zebrafish larvae. (A–C) Larvae were exposed to 10 µM cyA and 0.1% DMSO at 3 dpf for 3 days. (D–H) Embryos were injected with SHHMO, Gli2MO and Gli2Mis at the 1–4 cell stage; at 6 dpf, larvae were collected for assays . GAPDH served as a loading control. The means and s.d. were derived from triple duplicates. *P<0.05, **P<0.01, ***P<0.001 versus the WT groups or DMSO control groups (one-way ANOVA). The data were from three independent experiments. (A,D) SHH protein was detected by western blot assay. (B,E,G) Transcriptional level of autophagy-related genes was tested by qPCR assays. (C,F,H) Autophagy-related proteins LC3B II, ATG5 and P62 level were determined by western blot. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30992325), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

FASN Regulates HIF-alpha Protein Stability and HIF Target Gene Expression. (A) FASN siRNA downregulates HIF-alpha protein levels. PREC, HeLa and HCT116 cells were treated with FASN (+) or control (−) siRNA and harvested after 72 h. Cells were exposed to hypoxia (1% oxygen) or maintained in normoxia (21% oxygen) 4 h before harvesting and subjecting WCEs to western blot analysis. Unmodified (250 kDa) and modified (larger) FASN bands are indicated by large arrowheads, REDD1 by a small arrowhead, and HIF-1 alpha and HIF-2 alpha by arrows. (B) Downregulation of HIF-1 alpha and HIF-2 alpha in HCT116 cell clones carrying the FASN gene mutagenized using CRISPR-Cas9 (See Supplementary Fig. S3 for genomic DNA information). Protein samples were prepared in normoxia (left column) or hypoxia (right column) for western blot analysis. Asterisk indicates a non-specific band. (C) Quantification of HIF-target gene expression by qPCR upon downregulation of FASN in PREC. Samples were prepared from cells treated as described for panel A but with 24 h hypoxia. siRNAs were transfected in triplicates and mRNAs of the HIF target genes were measured in biological triplicates and technical duplicates as a total of 6 samples by qPCR. Values were normalized against the PPIA reference gene and control samples. (D) Loss of downregulation of HIF-2 alpha levels in ccRCC cells. pVHL-deficient 786-O and A498 cells were tested in FASN siRNA knockdown experiments. (E) Restoration of HIF-alpha regulation by stable expression of exogenous pVHL in SKRC10 cells. pVHL-WT or pVHL-Y112H were introduced into pVHL-deficient SKRC10 ccRCCs using a lentiviral vector carrying a weak promoter and HIF-alpha protein levels were examined. Specific and non-specific bands are indicated by arrows and an asterisk, respectively. (F) FASN and the E3 ligase components are present in the pVHL-containing complexes in a mutually exclusive manner. The indicated constructs were transfected into 293 T cells and WCEs prepared after 48 h were tested for protein-protein interactions by precipitation with an anti-Flag antibody-conjugated resin. Note that FASN failed to precipitate CUL2 and Elongin C, as shown in lanes 6 and 8. Experiments were repeated twice (D,E,F) or more (A,B), and the results were reproduced. The qPCR experiment (C) was performed once with biological triplicates. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28775317), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Etv5b activates the smn promoter leading to increased Smn levels in motoneurons.(a–d) Whole-mount in situ hybridization of etv5b expression in 24 hpf embryos. Expression pattern in the entire embryo (a), in magnified lateral (b) and dorsal (c) views as well as in a transverse section (d) of the trunk. In the latter case, yellow arrowhead denote signal in the motoneuron region. Scale bars: 100 μm (a) and 50 μm (b–d). nc, notochord. (e) RT-PCR in control (C - only psmn:mCherry) versus etv5b overexpressing (OE - etv5b mRNA and psmn:mCherry) embryos at early gastrula stage. (f,g) scIF of control (f) and etv5b MO (g) cells from pmnx1:eGFP embryos. H2B, GFP and Smn signals are shown in Z-projected confocal sections. GFP+ cells marked by white rectangles are magnified on the right. Scale bars: 10 μm for low and 5 μm for high magnification. (h) Relative Smn signal in control and in etv5b MO cells from one representative experiment. Exact values are (mean ± SD): 0.99 ± 0.25 (control) and 0.7 ± 0.14 (etvb5 MO), p = 0.003 with Wilcoxon Sum Rank Test. (i) Average relative Smn levels in control and in etv5b MO motoneurons. The exact value of decrease is 0.75 ± 0.04 (mean ± SD). N = 3 experiments, n = number of analyzed cells. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/srep27470), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

The A2AR and baicalin attenuated CXCR4 expression in the hypoxia-induced PAH mouse model. CXCR4 and MYH11 expression levels in mouse PASMCs were analyzed by immunofluorescence staining (a; n = 3). CXCR4 protein is stained red, and MYH11 is stained green to indicate the PASMCs (×400; scale bars indicate 50 μm). CXCR4 fluorescence intensity was calculated (b; n = 3). CXCR4 protein expression levels in lung tissue were examined by western blot. GAPDH served as an internal control (c, d; n = 3). Data are presented as the mean ± SD. #Value significantly greater than the corresponding value in saline-treated normoxic mice (##P < 0.01). *Value significantly less than the corresponding value in hypoxic mice (*P < 0.05, **P < 0.01). §Value significantly less than the corresponding value in baicalin-treated mice (§P < 0.05, §§P < 0.01). +Value significantly different between WT and A2AR−/− mice (+P < 0.05, ++P < 0.01). WTH, wild-type hypoxic; A2AR−/−H, A2AR−/− hypoxic; s, saline-treated Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28774332), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Bv, Ca, Gp, Pm, Xp, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Bv, Ca, Gp, Pm, Xp, 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	NB100-134AF350
Alexa Fluor 405	NB100-134AF405
Alexa Fluor 488	NB100-134AF488
Alexa Fluor 532	NB100-134AF532
Alexa Fluor 594	NB100-134AF594
Alexa Fluor 647	NB100-134AF647
Alexa Fluor 700	NB100-134AF700
Alexa Fluor 750	NB100-134AF750
Allophycocyanin	NB100-134APC
Biotin	NB100-134B
DyLight 350	NB100-134UV
DyLight 405	NB100-134V
DyLight 488	NB100-134G
DyLight 550	NB100-134R
DyLight 594	NB100-134DL594
DyLight 650	NB100-134C
DyLight 680	NB100-134FR
DyLight 755	NB100-134IR
FITC	NB100-134F
HRP	NB100-134H
Janelia Fluor 549	NB100-134JF549
Janelia Fluor 646	NB100-134JF646
PE	NB100-134PE
PerCP	NB100-134PCP
mFluor Violet 450 SE	NB100-134MFV450
mFluor Violet 500 SE	NB100-134MFV500
mFluor Violet 610 SE	NB100-134MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-134SS

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NB100-182

FANCD2 Antibody - BSA Free

(8 Reviews)

(215 Publications)

0.05 ml

NB100-182

<b>Biological Strategies Validation. </b>Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - AMPK-activating AICAR treatment activates FANCD2, a pivotal molecule of Fanconi anemia DNA damage signaling pathway. AICAR treatment induces FANCD2 monoubiquitination in transformed normal fibroblasts (GM00637I). GM00637I cells were treated with 1 mm 2-deoxyglucose, 0.25 mM AICAR, or 1 mm phenformin for 24 h. Lysates were subjected to western blotting with FANCD2 Antibody, phospho-AMPKalpha1 (T172), and AMPKalpha and beta-actin. In FANCD2 blots, the position of monoubiquitinated FANCD2 (Ub-FANCD2) is indicated by an arrow. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1002/2211-5463.12185) licensed under a CC-BY license.

Simple Western: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - Simple Western lane view shows a specific band for FANCD2 in 0.1 mg/ml of HeLa lysate using FANCD2 Antibody. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Immunocytochemistry/Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - Analysis using the Biotin conjugate of FANCD2 Antibody. FANCD2 colocalizes in vivo with another protein in SiHa cells after cell exposure to IR. Proliferating SiHa cells were exposed to 10 Gy of IR and double color immunofluorescence staining was performed after 8 h. Images were captured in a Kodak digital image system on a Leica fluorescence microscope.

<b>Biological Strategies Validation. </b>Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - AICAR-induced FANCD2 monoubiquitination is dependent on AMPK. Inhibitor of AMPK blocks AICAR-induced FANCD2 monoubiquitination in GM00637I normal fibroblasts. Cells were pretreated with 5 um of Compound C (an AMPK inhibitor) 1 h before treatment with 0.25 mm AICAR for 24 h. Cell lysates were subjected to immunoblotting with FANCD2 Antibody to visualize monoubiquitinated FANCD2 (Ub-ANCD2). Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1002/2211-5463.12185) licensed under a CC-BY license.

Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - The FA-BRCA Pathway Is Activated on the Sex Chromosomes during Meiosis. Schematic of the FA-BRCA pathway. FA proteins analyzed in this study are shown in color. Western blot analysis with three independent anti-FANCD2 antibodies (G33, E33, and Novus NB100-182 antibody: NB). K561R, PD20 cells expressing a mutated form of FANCD2 incapable of monoubiquitination; WT, PD20 cells complemented with wild-type FANCD2; Vector, PD20 cells containing empty vector. Image collected and cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S2211124716313298), licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - HES1K109E mutation does not impair MMC-induced HES1 foci formation. HeLa cells transfected with HA-tagged HES1 or HES1K109E coding vectors were processed for immunofluorescence 16 h following treatment with MMC (120 ng/ml). Cells were double-stained with anti-HA and anti-FANCD2 antibodies (mouse anti-HA at 1:500; rabbit anti-FANCD2, Novus Biologicals NB100-182 at 1:2000) followed with secondary antibodies (Goat anti-mouse Alexafluor-555, ThermoScientific A-32727 and goat anti-rabbit Alexafluor-488, ThermoScientific, A-11008). Cell nuclei were labelled with TO-PRO-3 Iodide stain (1:400, ThermoScientific, T3605). Cells were visualized at 100x magnification. Image collected and cropped by CiteAb from the following publication (//bmcresnotes.biomedcentral.com/articles/10.1186/s13104-018-3243-7), licensed under a CC-BY license.

Immunohistochemistry: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - The tissue sections were incubated with a primary antibody cocktail of FANCD2 Antibody (NB100-182) at a dilution of 1:1000 & a monoclonal anti-Ki67 mouse antibody at a dilution of 1:150 for 1 h at room temperature. Then the slides were incubated with a secondary antibody cocktail containing FITC conjugated anti-rabbit IgG and Alexafluor 594 donkey anti-mouse secondary. The sections were mounted on glass slides using a DAPI-containing embedding medium (Vysis Dapi 1, Abbott Laboratories, Downers Grove, IL, USA), then analyzed under a fluorescence microscope. FANCD2 (top) foci positive NSCL tumor, and FANCD2 (bottom)foci negative NSCL tumor. Magnification: 1000x. Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fonc.2014.00368/abstract), licensed under a CC-BY license.

Western Blot: FANCD2 Antibody [NB100-182] - Analysis of FANCD2 (Molecular weight: 164.1 KDa) using the HRP conjugate of FANCD2 Antibody (lot C) in HeLa WCE.

Western Blot: FANCD2 Antibody [NB100-182] - Functional assessment of Fanconi anemia pathway. 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. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1002/mgg3.350), licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: FANCD2 Antibody [NB100-182] - Mouse Embryonic Fibroblasts Untreated and treated with Mitomycin C for 24hr with FANCD2 Antibody diluted 1:500 in 15%BCS+0.3% Triton X 100 in PBS. This image was submitted via customer Review.

Immunohistochemistry: FANCD2 Antibody [NB100-182] - Staining of human prostate, glandular epithelium using FANCD2 Antibody. Image using the Biotin format of this antibody.

Representative images of the cellular response to RS (induced by a low dose of hydroxyurea; HU) in U2OS cells, as determined by IF staining for gamma H2AX and FANCD2A selected cell is defined by the yellow arrow, and is enlarged in the bottom panel. Scale bars are indicated.

Chemical library screening for small molecules that inhibit the Fanconi anemia pathway. (A) Schematic of the screening for small molecules that inhibit IR-induced FANCD2 foci formation. (B) Representative photomicrographs of EGFP-FANCD2 foci in PD20F-EGFP-FANCD2 cells untreated and treated with the indicated compounds at the indicated concentration. The cells were treated with compounds immediately before irradiation (15 Gy), and fixed after 12 hours. (C) FANCD2 foci in HeLa cells untreated and treated with the indicated compounds at the indicated concentration. The cells were fixed 8 hours after irradiation (10 Gy) and immunostained with anti-FANCD2 antibody. Scale bar = 20 um. Image collected and cropped by CiteAb from the following publication (//molecular-cancer.biomedcentral.com/articles/10.1186/1476-4598-11-26), licensed under a CC-BY licence.

The FA-BRCA Pathway Is Activated on the Sex Chromosomes during Meiosis. (F) Western blot analysis with three independent anti-FANCD2 antibodies (G33, E33, and Novus NB100-182 antibody: NB). K561R, PD20 cells expressing a mutated form of FANCD2 incapable of monoubiquitination; WT, PD20 cells complemented with wild-type FANCD2; Vector, PD20 cells containing empty vector. Image collected and cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S2211124716313298), licensed under a CC-BY licence.

HES1 mono-ubiquitination is not required for foci formation. a HES1K109E mutation does not impair MMC-induced HES1 foci formation. HeLa cells transfected with HA-tagged HES1 or HES1K109E coding vectors were processed for immunofluorescence 16 h following treatment with MMC (120 ng/ml). Cells were double-stained with anti-HA and anti-FANCD2 antibodies (mouse anti-HA at 1:500; rabbit anti-FANCD2, Novus Biologicals NB100-182 at 1:2000) followed with secondary antibodies (Goat anti-mouse Alexafluor-555, ThermoScientific A-32727 and goat anti-rabbit Alexafluor-488, ThermoScientific, A-11008). Cell nuclei were labelled with TO-PRO-3 Iodide stain (1:400, ThermoScientific, T3605). Cells were visualized at 100x magnification. b HES1 lysine 109 plays a role in MMC-induced activation of the HES1 promoter. COS-1 cells transfected with pHES1pro-Luc and HES1 or HES1K109E coding vectors (0.034 ug) were treated with 120 ng/ml MMC. Empty vectors were used as controls. Experiments were done twice in triplicates. 2-way ANOVA with Bonferroni correction test, *p 10 larvae/sample). The blots were probed with antibodies against Lc3 and Actin as a loading control and Lc3-II/Lc3-I ratios are indicated below. Western blots were repeated twice with independent extracts. 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.

Ddx19 is specifically required for the nuclear localization of MKL1.(a) Western blotting (WB) of cells treated with control (Ctrl) or Ddx19 siRNAs (si). (b) Localization of MKL1-GFP (left panels) and endogenous MKL1 ( alpha -MKL1; right panel) in serum (FBS)-stimulated NIH 3T3 cells. Scale bars, 20 μm. (c) Quantification of MKL1-GFP localization in serum-starved (0.3% FBS), serum-stimulated (15% FBS) and LMB-treated control (−) or Ddx19 (+) siRNA-transfected cells. C, cytoplasmic; N/C, pancellular; N, nuclear. One hundred cells per point; n=3; error bars are s.e.m. (d) Localization of nuclear proteins in NIH 3T3 cells transfected with control (−) or Ddx19 (+) siRNA. SRF (serum response factor), RanBPI-delta NES (Ran binding protein 1 lacking nuclear export signal), hnRNPU (heterogeneous nuclear ribonucleoprotein U), Smad4+LMB (Smad4 stimulated with leptomycin B), MC (myocardin). More than fifty cells per point; n=2; error bars are s.d. (e) Localization of NLS-2GFP-NES in serum-starved (0.3% FBS) or LMB-treated cells transfected with control (−) or Ddx19 (+) siRNAs. One hundred cells per point; n=2; error bars are s.d. 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, Av, Ca, Ma, Pm, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Av, Ca, Ma, Pm, 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 405	NB100-182AF405
Alexa Fluor 488	NB100-182AF488
Alexa Fluor 647	NB100-182AF647
Alexa Fluor 700	NB100-182AF700
Biotin	NB100-182B
DyLight 405	NB100-182V
DyLight 488	NB100-182G
DyLight 550	NB100-182R
DyLight 650	NB100-182C
FITC	NB100-182F
HRP	NB100-182H

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-182SS

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NB400-135

CHREBP Antibody - BSA Free

(7 Reviews)

(145 Publications)

0.1 ml

NB400-135

Immunohistochemistry-Paraffin: CHREBP Antibody - BSA Free [NB400-135] - CHREBP Antibody [NB400-135] - Staining of human colon, epithelium at a 40X magnification.

Chromatin Immunoprecipitation: CHREBP Antibody - BSA Free [NB400-135] - CHREBP Antibody [NB400-135] - Effects of glucose on ChREBP binding.HepG2 cells were treated with low (2.7 mM) and high (25 mM) glucose for 8 h. Chromatin was isolated and fragmented, and ChIP was performed with control IgG or anti-ChREBP antibody. Validated primers for each gene were used for quantitative real-time PCR. The data presented as fold increase for the signal from anti-ChREBP relative to control IgG. The negative control, Cyclo, showed no enrichment (data not shown). Values represent the mean +/- S.D. of three independent samples. *p<0.005 vs. IgG, #p<0.0001 vs. 2.7 mM glucose with anti-ChREBP. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0022544), licensed under a CC-BY license.

Western Blot: CHREBP Antibody - BSA Free [NB400-135] - CHREBP Antibody [NB400-135] - Detection of ChREBP in 20 ug of human hepatocyte lysate using NB400-135. 5-10 second film exposure.

Western Blot: CHREBP Antibody - BSA Free [NB400-135] - CHREBP Antibody [NB400-135] - Detection of ChREBP in liver nuclear extracts from well-fed rats. 7% SDS-PAGE gel, 1:1000 dilution of NB400-135. Photo courtesy of Dr. Uyeda, UT Southwestern University.

Western Blot: CHREBP Antibody - BSA Free [NB400-135] - CHREBP Antibody [NB400-135] - RetSat depletion in mouse liver reduces protein levels and target gene expression of ChREBP. RetSat and ChREBP protein expression by immunoblotting. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-017-00430-w), licensed under a CC-BY license.

Western Blot: CHREBP Antibody - BSA Free [NB400-135] - CHREBP Antibody [NB400-135] - Protein expression levels of carbohydrate-responsive element-binding protein (ChREBP), in livers of rats receiving normal water (CON), a 13% (w/v) glucose solution (GLU), a 13% (w/v) fructose solution (FRUC), or a 0.4% (w/v) aspartame solution (n = 6 per diet group). All data were normalized to beta-actin expression levels and are expressed relative to the controls (CON). Data are expressed as means +/- SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. CON; +p < 0.05, ++p < 0.01, +++p < 0.001 vs. ASP; p < 0.01, p < 0.001 vs. GLU. Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/2072-6643/9/5/476), licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: CHREBP Antibody - BSA Free [NB400-135] - CHREBP Antibody [NB400-135] - HeLa cells were fixed for 10 minutes using 10% formalin and permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-ChREBP (NB400-135) at a 1:200 dilution overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:500 dilution. Alpha tubulin was used as a co-stained at 1:1000 and detected with an anti-mouse DyLight 550 (Red) at a 1:500. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry: CHREBP Antibody - BSA Free [NB400-135] - CHREBP Antibody [NB400-135] - Analysis of CHREBP in mouse liver using DAB with hematoxylin counterstain.

<b>Genetic Strategies Validation. </b>Knockout Validated: CHREBP Antibody - BSA Free [NB400-135] - Immunoblot analysis of ChREBP in liver lysates of control and L-Chrebp -/- mice. Aliquots (60 ug of protein) of liver whole-cell lysates were subjected to SDS-PAGE and immunoblot analysis with anti-ChREBP and anti-calnexin antibodies. ChREBP delta denotes a truncated aberrant ChREBP protein present only in lysates prepared from L-Chrebp -/- livers. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29335275/) licensed under a CC-BY license.

Analysis of protein levels in livers transfected with GK and GKA456V. Livers excised from mice in postabsorptive state were homogenized and resolved by Western blot. (a, b) Representative blots from three independent experiments. The densitometric analysis is presented (N = 4, *P < 0.05 versus pControl, ***P < 0.001 versus pGKA456V). (c) Liver sections of 5-hour fasted mice injected with pControl, pGK, or pGKA456V were immunostained with Glc6Pase antibody. TO-PRO-3 was used to visualize nuclei.

LXR mediate the induction of lipogenesis induced by an oleic acid-rich diet. (E) Cytoplasmic and nuclear expression levels of LXR, SREBP-1c and ChREBP assayed by Western Blotting. Data are the mean+/-SEM of values measured in LXR+/+ and LXR-/- mice fed REF or OLIV diet. a Significant genotype effect. b Significant difference versus REF diet (n = 6 mice per group). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28732092), licensed under a CC-BY licence.

ChREBP knockdown inhibited glycolysis, lipogenesis and p21 in HT29 cells. (B) Western-blot of ChREBP, phospho-p53 and p21 and their quantification on the right. beta-actin was served as a loading control. Proteins were extracted from cells transfected with sicontrol and siChREBP after 48 hours. The quantification of western blot was normalized to beta-actin. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32144313), licensed under a CC-BY licence.

Effects of glucose on ChREBP binding.HepG2 cells were treated with low (2.7 mM) and high (25 mM) glucose for 8 h. Chromatin was isolated and fragmented, and ChIP was performed with control IgG or anti-ChREBP antibody. Validated primers for each gene were used for quantitative real-time PCR. The data presented as fold increase for the signal from anti-ChREBP relative to control IgG. The negative control, Cyclo, showed no enrichment (data not shown). Values represent the mean +/- S.D. of three independent samples. *p<0.005 vs. IgG, #p<0.0001 vs. 2.7 mM glucose with anti-ChREBP. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/21811631), licensed under a CC-BY licence.

Sucrose intake increases hepatic triglyceride content via sex-specific mechanisms. e) Representative western blots demonstrating that the abundance of proteins encoded by the transcripts differentially expressed (as well as ChREBP and its target PKLR) between males and female mice in response to sucrose intake. Statistics were derived following densitometric quantification of bands from samples run on a single gel that were normalized to total protein and then log transformed and compared using two-way ANOVA with post hoc Tukey's tests (FAS: ap= 1.43-4, bp = 1.55-4, dp = 0.044, ep = 2.80-6; pACC: ap = 0.001, bp = 0.007, ep = 2.8-4, fp = 0.041; ACC: ap = 0.001, bp = 0.003, ep = 2.99-4, fp = 0.014; ChREBP: ep = 0.024, fp = 0.010; MC: n = 6, MS: n = 6, FC: n = 6, FS: n = 6). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/36229459), licensed under a CC-BY licence.

FXR inhibits glucose-induced proglucagon expression. (c) ChREBP and FXR western-blots after FXR immunoprecipitation on lysates from cytoplasm and nucleus of GLUTag cells treated or not with GW4064 (5 umol L-1) in presence or not of glucose (5.6 mmol L-1) (performed 2 times). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms8629), licensed under a CC-BY licence.

Raffinose stimulates transcription of genes involved in epidermal barrier function in HaCaT cells.(a and b) HaCaT cells were treated with vehicle, 1 uM or 10 uM raffinose (Raf), or 1 uM TO901317 (T17) for 24 h. Expressions of transcripts (a) and proteins (b) were analyzed by qRT-PCR or western blotting, respectively. FLG; filaggrin. IVL; involucrin, LOR; loricrin, AQP3; aquaporin3. (c) HaCaT cells were transfected with siGFP control, siLXRalpha, or siLXRbeta, and then treated with 1 uM raffinose for 24 h. Expression of proteins was analyzed by western blotting. The original blots are shown in Supplementary Fig. S9. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28266648), licensed under a CC-BY licence.

Hepatic expression of mRNAs encoding carbohydrate response element-binding protein (ChREBP) (a), liver pyruvate kinase (LPK) (c), and microsomal triglyceride transfer protein (MTTP) (d), and nuclear ChREBP protein (b) in water-control, 10% fructose solution-control, and fructose pair-fed oleanolic acid- (OA-) treated rats at week 10. Animals were administered with OA (25 mg/kg/day) or vehicle (OA: 0 mg/kg, 5% Gum Arabic) by oral gavage daily for 10 weeks. mRNA was determined by real-time PCR. Protein expression was determined by Western blot. Data are means +/- SEM (n = 6 each group). *P < 0.05. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23737835), licensed under a CC-BY licence.

Protein expression levels of (D) carbohydrate-responsive element-binding protein (ChREBP); (E) 128 kDa precursor sterol regulatory element-binding protein-1c (SREBP-1c); and (F) 65 kDa cleaved SREBP-1c, in livers of rats receiving normal water (CON), a 13% (w/v) glucose solution (GLU), a 13% (w/v) fructose solution (FRUC), or a 0.4% (w/v) aspartame solution (n = 6 per diet group). All data were normalized to beta-actin expression levels and are expressed relative to the controls (CON). Data are expressed as means +/- SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. CON; +p < 0.05, ++p < 0.01, +++p < 0.001 vs. ASP; **p < 0.01, ***p < 0.001 vs. GLU. Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/2072-6643/9/5/476), licensed under a CC-BY licence.

Optical redox ratio (ORR) demonstrates reduced glycolytic phenotype following 24 hours of YC-1 treatment. ORR of A549 and A549RR cells before, 12 and 24 hours following 50 μM YC-1 treatment to inhibit HIF-1 (a,b). Immunoblotting for HIF-1 alpha (c) and PDK-1 (d) before and 24 hours following YC-1 treatment to verify in of HIF-1 alpha . Representative immunoblots were horizontally cropped at the molecular weight indicated (e). Uncropped exposures can be seen in Supplemental Fig. 1. Experiments were performed in triplicate across 3 independent experiments. *p < 0.05 compared to A549 0 hr control, †p < 0.05 compared to A549RR 0hrs. Scale bar in images represents 50 μm. ME – Main effect of cell type and time; however, no significant interactions were found between the two main effects. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/s41598-018-27262-y), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Excessive AP-endonuclease activity in cells with p53 impairment leads to accumulation of genomic instability. (A) Representative western blotting analysis on WI38 (SV40) cells shows rescue of gamma H2AX staining upon co-depletion of XRCC1 and APE1. (B) Neutral Comet assay on WI38 (SV40) fibroblasts shows accumulation of DSBs upon XRCC1-depletion and rescue after co-depletion of XRCC1 and APE1. Results are expressed as mean ± SD of three independent experiments. (C) Representative western blotting analysis on WI38 (SV40) cells shows rescue of gamma H2AX staining after depletion of XRCC1 in presence of APE1 inhibitor III (APE1i III, 5 μM for 24 h), or AR03 (2.5 μM for 24 h). Actin was used as loading control in all western blotting experiments. Image collected and cropped by CiteAb from the following open publication (//academic.oup.com/nar/article-lookup/doi/10.1093/nar/gkw015), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Effects of resveratrol on the expression of SIRT1, PGC-1 alpha , HIF-1 alpha and apoptosis proteins in vitro. (A) The expression of SIRT1, PGC-1 alpha , and Bcl-2 proteins were lower in iohexol treated HK-2 cells compared with control cells, which was reversed by treatment with resveratrol. The expression of HIF-1 alpha , Bax, cleaved caspase-3, and Cyt-C were markedly increased after iohexol injection, which was reversed by treatment with resveratrol. Furthermore, this effect was enhanced by HIF-1 alpha inhibitor, 2-MeOE2. (B–H) Relative densitometry of SIRT1–PGC-1 alpha –HIF-1 alpha signaling proteins expressed as mean ± standard error. *P < 0.05 vs. Cont; #P < 0.05 vs. PC–AKI. Cont, control; Res, resveratrol; PC–AKI, post-contrast acute kidney injury; SIRT1, silent information regulator l; HIF-1 alpha , hypoxia-inducible transcription factor-1 alpha ; Cyt-C, cytochrome C; PGC-1 alpha , peroxisome proliferator-activated receptor gamma coactivator-1 alpha; Ex527, SIRT1 specific inhibitor; LG, low glucose. Groups: Res, HG+res+iohexol; Ex527, HG+Ex527+iohexol; PC–AKI, HG+iohexol; 2-MeOE2, HG+res+2-MeOE2+iohexol; Cont, LG. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31402864), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

EpEX and EpCAM activate a STAT3-HIF2 alpha signal for EpEX/EpCAM-mediated iPSC formation.(A) iPSCs were infected with two different EpCAM shRNAs (two clones, #1 and #2). The protein expressions of EpCAM and HIF2 alpha were detected by Western blotting. (B) MEFs were stimulated by EpEX (1 μg/mL) at the indicated times. Nuclear-translocation was detected with a specific antibody against HIF2 alpha (n = 3). (C) Immunofluorescence staining was performed to detect subcellular localization of HIF2 alpha . Nuclei were stained with DAPI. Scale bar: 10 μm. (D) MEFs were treated with STAT3 inhibitor (WP1066, 10 μM), and then stimulated with EpEX for 30 min. The nuclear-translocation of HIF2 alpha was detected by Western blotting with anti-HIF2 alpha antibody (n = 3). (E) iPSC morphology was observed at day 20 after induction. Reprogramming of Oct4-GFP MEFs was induced by transfection of OSKM, OE + EpEX, and KE + EpEX with or without STAT3 inhibitor WP1066, or HIF2 alpha shRNA (n = 3). Scale bar: 50 μm. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/srep41852), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Cell cycle block under PHD3 depletion is accompanied by p27 induction. a PHD3 depletion induces a cell cycle block in G0/G1. HeLa and renal cell adenocarcinoma cells (786-O) were transfected with control (siScr) or PHD3 targeted (siPHD3) siRNA followed by synchronization at G0 and 24-h hypoxic exposure. Cell cycle progression was monitored by FACS analysis 8 h after cell cycle release. The combined means of three independent experiments are presented (±SEM) shown in the tables below. b PHD3 depletion induces p27 expression in HeLa cells and in 786-O cells under hypoxia (1 % O2) and normoxia (21 % O2) by siPHD3 and independent adenoviral shRNA against PHD3. p21 or p16 expression is not elevated by PHD3 knockdown. c Depletion of either PHD1 or PHD2 by siRNA does not increase p27 expression in 786-O cells Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/26223520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

GAD65/67-positive neurons expressed HIF-1 alpha under hypoxic conditions. A) HIF-1 alpha expression co-localized with GAD65/67-ir in neurons exposed to hypoxia when compared to normoxia (upper panel) or GAD65/67-negative neurons in hypoxia (bottom panel, open arrow). B) Quantification shows the percentage of HIF-1 alpha -expressing GAD65/67-positive neurons after hypoxia in vitro (mean ± SD; from n = 6 cultures). C)In vivo immunostaining illustrates HIF-1 alpha -positive (bottom panel, solid arrow) and HIF-1 alpha -negative (bottom panel, open arrow) in GAD65/67-ir neurons in the ipsilateral region, whereas the contralateral region shows no HIF-1 alpha staining in GAD65/67-ir neurons (see Figure 1 for region selection). Scale bars, 10 μm (A); 20 μm (B). Image collected and cropped by CiteAb from the following open publication (//actaneurocomms.biomedcentral.com/articles/10.1186/2051-5960-2-51), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Absence of Zizimin proteins, but normal numbers of hematopoietic cells in KO mice. (A-B) Western blotting for Ziz2 (A) or Ziz3 (B) Protein lysates (30 μg/lane) were loaded and designated antibodies were reacted with the membranes. Replicated membranes were used for all antibodies. Zizimin proteins were absent in KO mice (8 weeks old, male). (C-F) The numbers of hematopoietic cells were counted after the hemolytic reaction (C-E) or flow cytometric analysis (F). No significant difference was observed among the groups (C-F). Three mice (8 weeks old, female) per group from three independent experiments (one mouse per group per experiment) were used. 2KO: Ziz2 KO. 3KO: Ziz3 KO. Image collected and cropped by CiteAb from the following open publication (//immunityageing.biomedcentral.com/articles/10.1186/s12979-015-0028-x), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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 (A). Clusters of CNHC-UMF and -BF without reactivity for CAIX antibodies (B and C, respectively). A single CNHC-MF with positive cytoplasmic (D) and without staining for CAIX (E). Single CAIX-negative CNHC-UMF and -BF (F and G, respectively). Image collected and cropped by CiteAb from the following open publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-214), 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:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB400-135AF350
Alexa Fluor 405	NB400-135AF405
Alexa Fluor 488	NB400-135AF488
Alexa Fluor 532	NB400-135AF532
Alexa Fluor 594	NB400-135AF594
Alexa Fluor 647	NB400-135AF647
Alexa Fluor 700	NB400-135AF700
Alexa Fluor 750	NB400-135AF750
Allophycocyanin	NB400-135APC
Biotin	NB400-135B
DyLight 350	NB400-135UV
DyLight 405	NB400-135V
DyLight 488	NB400-135G
DyLight 550	NB400-135R
DyLight 594	NB400-135DL594
DyLight 650	NB400-135C
DyLight 680	NB400-135FR
DyLight 755	NB400-135IR
FITC	NB400-135F
HRP	NB400-135H
Janelia Fluor 549	NB400-135JF549
Janelia Fluor 646	NB400-135JF646
PE	NB400-135PE
PerCP	NB400-135PCP
mFluor Violet 450 SE	NB400-135MFV450
mFluor Violet 500 SE	NB400-135MFV500
mFluor Violet 610 SE	NB400-135MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB400-135SS

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NB110-57130

TRF-2 Antibody - BSA Free

(2 Reviews)

(136 Publications)

0.1 ml

NB110-57130

Western Blot: TRF-2 Antibody [NB110-57130] - Analysis of HeLa whole cell lysate (A), HeLa nuclear cell lysate (B), k562 cell lysate (C), HepG2 cell lysate (D), NIH/3T3 cell lysate (E), CHO cell lysate (F), PC12 cell lysate (G), and Cos7 cell lysate (H) using antibody at a concentration of 2 ug/mL.

Immunohistochemistry-Paraffin: TRF-2 Antibody [NB110-57130] - Analysis in xenografted human breast cancer tissue using DAB with hematoxylin counterstain.

Immunohistochemistry-Paraffin: TRF-2 Antibody [NB110-57130] - Analysis of FFPE human breast cancer tissue with rabbit polyclonal TRF2 antibody at a dilution of 1:200. The staining was developed with HRP-DAB detection method and the counterstaining was performed using hematoxylin. This TRF2 antibody generated an expected nuclear signal in all the cancer cells and the stromal cells. In the tested section, only a subset of myoepithelial cells showed positivity for this protein.

Immunocytochemistry/Immunofluorescence: TRF-2 Antibody [NB110-57130] - 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-TRF-2 Antibody NB110-57130 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.

Flow Cytometry: TRF-2 Antibody [NB110-57130] - An intracellular stain was performed on HeLa cells with TRF-2 Antibody NB110-57130 (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).

Chromatin Immunoprecipitation: TRF-2 Antibody [NB110-57130] - Analysis in mouse. Titrated TRF2 antibody to determine concentration required for ChIP experiment. ChIP image submitted by a verified customer review.

<b>Genetic Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: TRF-2 Antibody [NB110-57130] - RNAi-mediated depletion of human separase (ESPL1) induces TIFs. Control scrambled siRNA- (control) and ESPL1 siRNA-treated fibroblasts stained with anti-p53-binding protein 1 (53BP1; green) and anti-TRF2 (red). It is noteworthy that in ESPL1 siRNA-treated cells, 53BP1 signals frequently overlap with TRF2 signals marking the TIFs. Scale bar, 5 um. Image collected and cropped by CiteAb from the following publication (//www.nature.com/doifinder/10.1038/ncomms10405), licensed under a CC-BY license.

Flow Cytometry: TRF-2 Antibody [NB110-57130] - An intracellular stain was performed on HeLa cells with TRF-2 Antibody NB110-57130AF488 (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 488.

Telomere deprotection contributes to replication stress lethality. A) Western blots of whole cell extracts from HT1080 6TG cells stably transduced with control, TRF2 shRNA (TRF sh-F) or TRF2 over expression (TRF2OE) vectors. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31530811), licensed under a CC-BY licence.

RNAi-mediated depletion of human separase (ESPL1) induces TIFs.(a) Control scrambled siRNA- (control) and ESPL1 siRNA-treated fibroblasts stained with anti-p53-binding protein 1 (53BP1; green) and anti-TRF2 (red). It is noteworthy that in ESPL1 siRNA-treated cells, 53BP1 signals frequently overlap with TRF2 signals marking the TIFs. Scale bar, 5 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26778495), licensed under a CC-BY licence.

$Polyphyllin I triggers PINK1/PARK2-dependent mitophagyA. MDA-MB-231 cells were treated with 8 μM PPI for different periods of time as indicated, and PARK2, P62, and ubiquitin (UB) levels in mitochondrial fractions were determined by western blot. B. Cells were cotransfected with GFP-UB and RFP-mito and treated with 8 μM PPI for 9 h, after which PARK2 (Alexa Fluor 405, pink) and P62 (Alexa Fluor 647, blue) immunostaining was detected using confocal microscopy. C. Cells cotransfected with GFP-UB and RFP-LC3 were treated with 8 μM PPI for 9 h, after which PARK2 (Alexa Fluor 405, pink) and TOMM20 (Alexa Fluor 647, blue) immunostaining was detected using confocal microscopy. Scale bars: 10 μm. D-E. MDA-MB-231 cells were treated with 8 μM PPI for different periods of time as indicated; whole-cell lysates were then separated on 8% SDS-PAGE gels and analyzed by western blot using the anti-PINK1 antibody. Relative full-length (∼63 kDa) and cleaved (∼52 kDa) PINK1 levels were quantified by densitometry and normalized to Tubulin. The results were expressed as a percentage of control, which was set at 100%. Data are presented as mean ± SD (*P< 0.01 vs. the control). F. Cells were treated with 8 μM PPI for different periods of time as indicated, and whole-cell lysates were then subjected to western blot analysis. G. Cells were treated with 8 μM PPI for 9 h, after which mitochondrial fractions were prepared and subjected to immunoprecipitation using anti-PINK1 antibody; associated PARK2 was detected using immunoblotting. H. RFP-mito-expressing MDA-MB-231 cells were treated with 8 μM PPI for 9 h, and PINK1 (Alexa Fluor 488, green) and PARK2 (Alexa Fluor 405, pink) immunostaining were evaluated using confocal microscopy. Scale bars: 10 μm. Image collected and cropped by CiteAb from the following open publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.14413), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

PHD3 depletion stabilizes hypoxic p27 expression by increasing p27 half-life. a Cell cycle arrest at G0 and subsequent release shows an increase of p27 expression in siPHD3 exposed cells. b Quantification for p27 expression under PHD3 depletion at indicated time points after cell cycle release in HeLa and 786-O cells. Asterisk indicates significant difference (p < 0,05; n = 3). c Cell cycle arrest at G0 and inhibition of protein synthesis with cycloheximide indicate increased p27 stability in PHD3 depleted HeLa cells. d Quantification of p27 expression using siPHD3 or control at indicated time points. Four independent experiments (± SEM) are shown (p < 0,05; n = 4). e Analysis of p27 stability in 786–0 cells by cycloheximide chase during reoxygenation after 24 h hypoxia demonstrates markedly increased half-life of p27 upon PHD3 depletion Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/26223520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemical staining of HIF-1 alpha , VEGF-A and VEGF-C in normal renal tissue (A-C) and clear cell renal cell carcinoma (CCRCC) (D-F). A homogeneous cytoplasmic staining of tubular cells and weak staining in glomerules was observed with HIF-1 alpha (A), while VEGF-A and VEGF-C were positive in tubular cells, glomerular mesangium and interstitial macrophages (B and C). In CCRCC, HIF-1 alpha immmunoreactivity was nuclear and/or cytoplasmic (D), while it was perimembranous and/or diffuse cytoplasmic for VEGF-A and VEFG-C (E and F). (magnification ×200). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/19302703), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Resistance of FANCA-mutant cells with defective FANCD2 function to hydrogen peroxide (H2O2). (a) Representative images illustrating staining for subnuclear FANCD2 foci in isogenic fibroblast pairs, either deficient for FANCD2 (PD20) or FANCA (PD220) and their respective wild-type (wt) complemented counterparts. Foci were visualized three hours after treatment with H2O2 (25 μM for 2 hours). (b) Clonogenic survival of cells with or without wild-type FANCA after treatment with varying concentrations of H2O2. Datarepresent logarithmic means +/− standard error from 15-repeat experiments. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/18483568), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$gamma -Taxilin ablation causes ER stress responses and apoptosis in normoxic cells. (a) Knockdown of gamma -taxilin expression by small interfering RNA (siRNA) in HeLa S3 cells. Two different siRNA constructs are equally effective. HeLa S3 cells were treated with solvent alone (Mock), control siRNA (Co), or gamma -taxilin siRNA ( gamma -tax-1 or -2) for 96 h. (b) Induction of apoptosis by gamma -taxilin ablation in HeLa S3 cells. gamma -Taxilin ablation causes apoptosis in HeLa S3 cells, but Mock or Co treatment did not. Bar graph shows fractions of annexin-positive cells (means±S.D., n=3). *Significantly different from Mock- or Co treatment (P <0.001, Tukey–Kramer test). (c) Confocal microscopy demonstrates coincidence of gamma -taxilin depletion and apoptotic nuclei in HeLa S3 cells treated with gamma -taxilin siRNA. Scale bar, 10 μm. (d) gamma -Taxilin depletion triggers ER stress responses in HeLa S3 cells. Upper panels, taxilin and NAC proteins; middle panels, UPR proteins; and lower panel, beta -actin. (e) gamma -Taxilin ablation induces accumulation of ubiquitinated proteins in HeLa S3 cells. Cell lysates were analyzed on 7.5% SDS-PAGE, followed by immunoblotting with antibodies specific for anti-mono- and polyubiquitinated conjugates (upper panel) and beta -actin (lower panel) Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/25880086), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

$Damage response is restricted to MG. (A) RT-PCR analysis for Slc1a3, the gene encoding GLAST, at the indicated times after injury. (B) RT-PCR for MG and a photoreceptor-specific marker (Nrl) in GLAST-positive and negative fractions, after MACS in intact retinas. (C–F) qPCR quantification of Oct4, Nanog, Lin28, and Dnmt3b expression levels at the indicated time after injury in MACS GLAST-positive and negative fraction; C, intact retina as control (Student's t-test; ***p < 0.001; **p < 0.01; *p < 0.05). Image collected and cropped by CiteAb from the following open publication (//journal.frontiersin.org/article/10.3389/fnins.2016.00523/full), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Effect of ET-1 and Cx43 on HIF-1 alpha expression and glucose uptake when c-Src is inhibited.Astrocytes were preincubated with 100 ng/µL PP2 (c-Src inhibitor) or 100 ng/µL PP3 (inactive analogue) for 1 h. Then, cells were incubated in the absence (control) or presence of 0.1 µM ET-1 for 24 h. A) HIF-1 alpha Western blot and quantification. The results are expressed as percentages of the level found in the controls treated with PP3 and they show that the inhibitor of c-Src PP2 prevented the up-regulation of HIF-1 alpha promoted by ET-1. ***p<0.001 versus the absence of ET-1. B) Glucose uptake expressed as pmol of 2-deoxyglucose taken up per hour and per milligram of protein. The results show that the inhibitor of c-Src PP2 prevented the increase in the rate of glucose uptake promoted by ET-1. ***p<0.001 versus the absence of ET-1. C) Astrocytes were preincubated with 100 ng/µL PP2 or 100 ng/µL PP3 for 1 h. Then, cells were transfected with NT-siRNA or with Cx43-siRNA and after 48 h HIF-1 alpha was analysed by Western blot. The results are expressed as percentages of the level found in the PP3 NT-siRNA and they show that the inhibitor of c-Src PP2 prevented the up-regulation of HIF-1 alpha promoted by silencing Cx43. ***p<0.001 versus the corresponding NT-siRNA. Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.pone.0032448), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Genotyping of transgenes.PCR genotyping was performed for all four transgenes as described in materials and methods. Sizes of the amplified products obtained are: 240 bp for Hif-1 alpha (wild type); 274 bp for Hif-1 alpha flox/flox; 410 bp for Hif-2 alpha (wild type); 444bp for Hif-2 alpha flox/flox; 370 bpfor Cre transgene; 350 bpfor rtTA transgene. One representative sample was genotyped for the four transgenes from each of three generated mouse strains: SPC-rtTA-/tg/(tetO)7-Cre-/tg/Hif-1 alpha fl/fl (Lane 1), SPC-rtTA-/tg/(tetO)7-Cre-/tg/Hif-2 alpha fl/fl mouse (Lane 2), and SPC-rtTA-/tg/(tetO)7-Cre-/tg/Hif-1 alpha /2 alpha fl/fl (Lane 3)). Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.pone.0139270), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

The expression patterns of Yap and GFAP-Cre recombinase in postnatal mouse eyes. (A) Schematic of a transverse section of mouse eye. (B-C) Immunostaining with anti-Yap antibody (green) on frozen eye sections at different ages. Nuclei were counterstained with DAPI (blue). Yap staining was detected in scattered cells within the INL (arrowheads) and GCL of the retina and the lens epithelium (arrows). (D) Cre recombinase (red) was expressed in the lens epithelium and INL, GCL of retina in frozen eye sections of Tomatof/+; GFAP-Cre mice at P14. Nuclei were counterstained with DAPI (blue). LE, lens epithelium; TZ, transitional zone; RPE, retinal pigment epithelium; OS, outer segment; IS, inner segment; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer. Scale bars: 25 μm (B-C), 100 μm (D). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31011480), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Representative picture of western blot in histopathologically unchanged tissue (N) and primary cancerous tissue (C) from patients with CRC. Immunodetection of bands was performed with Rp anti- PHD1, - PHD2, - PHD3 and - FIH Ab, followed by incubation with goat anti-rabbit HRP-conjugated Ab. The membrane was stripped and incubated with Rp anti-GAPDH Ab, followed by incubation with goat anti-rabbit HRP-conjugated Ab. Bands were revealed using SuperSignal West Femto Chemiluminescent Substrate, Thermo Fisher Scientific (Rockford, IL) and Biospectrum® Imaging System 500, UVP Ltd. (Upland, CA). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/24195777), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, ChHa, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, ChHa, Pm

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB110-57130AF350
Alexa Fluor 405	NB110-57130AF405
Alexa Fluor 488	NB110-57130AF488
Alexa Fluor 532	NB110-57130AF532
Alexa Fluor 594	NB110-57130AF594
Alexa Fluor 647	NB110-57130AF647
Alexa Fluor 700	NB110-57130AF700
Alexa Fluor 750	NB110-57130AF750
Allophycocyanin	NB110-57130APC
Biotin	NB110-57130B
DyLight 350	NB110-57130UV
DyLight 405	NB110-57130V
DyLight 488	NB110-57130G
DyLight 550	NB110-57130R
DyLight 594	NB110-57130DL594
DyLight 650	NB110-57130C
DyLight 680	NB110-57130FR
DyLight 755	NB110-57130IR
FITC	NB110-57130F
HRP	NB110-57130H
Janelia Fluor 549	NB110-57130JF549
Janelia Fluor 646	NB110-57130JF646
PE	NB110-57130PE
PerCP	NB110-57130PCP
mFluor Violet 450 SE	NB110-57130MFV450
mFluor Violet 500 SE	NB110-57130MFV500
mFluor Violet 610 SE	NB110-57130MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB110-57130SS

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NB600-1252

Ki67/MKI67 Antibody (SP6) - Unpurified

(4 Reviews)

(90 Publications)

0.5 ml

NB600-1252

Immunohistochemistry-Paraffin: Ki67/MKI67 Antibody (SP6) - Unpurified [NB600-1252] - Formalin fixed paraffin embedded human tonsil stained with Ki-67 antibody.

Immunohistochemistry-Paraffin: Ki67/MKI67 Antibody (SP6) - Unpurified [NB600-1252] - Ki-67/MKI67 Antibody (SP6) [NB600-1252] - Formalin fixed paraffin embedded human tonsil stained with Ki-67 antibody.

Immunohistochemistry: Ki67/MKI67 Antibody (SP6) - Unpurified [NB600-1252] - Paraffin-embedded alcohol fixed rat spleen tissue (20x). Antigen retrieval pH 9. Ki67 dilution 1:100 incubation ON 4C. This image was submitted via customer review.

Immunohistochemistry: Ki67/MKI67 Antibody (SP6) - Unpurified [NB600-1252] - PW-12 is efficacious in a GEM model of MYCN-amplified neuroblastoma. Tumor-bearing TH-MYCN mice were treated for 14 days with either 25 mg/kg of PW-12 (n = 3) or vehicle alone (n = 3). Histology for Ki67 and c-Caspase 3 shows increase in apoptosis and decrease in proliferation in treated tumors. Image collected and cropped by CiteAb from the following publication (//www.frontiersin.org/Cancer_Molecular_Targets_and_Therapeutics/10.3389/fonc.2015.00111/abstract), licensed under a CC-BY license.

Immunohistochemistry: Ki67/MKI67 Antibody (SP6) - Unpurified [NB600-1252] - Ki-67/MKI67 Antibody (SP6) [NB600-1252] - Ki67 staining in mouse thyroid tissue at pre-tumor stage (green). Dilution is 1:100. This image was submitted via customer Review.

Immunohistochemistry-Paraffin: Ki67/MKI67 Antibody (SP6) - Unpurified [NB600-1252] - Ki-67 (NB600-1252) immunoreactivity in an FFPE section of mouse small intestine. Primary antibody was diluted 1:100 and left on sections for 1h at room temperature. Secondary antibody was Horse Anti-Rabbit HRP. Image from verified customer review.

Immunohistochemistry of tissue slices treated with trastuzumab and docetaxel. (Ki-67, arrows; A-C) and (cleaved caspase-3, arrows; D-F) were quantified by counting the number of positively stained cells in 4 fields of view at x40. Images are representative of untreated (A, D), trastuzumab at 100 µg/ml (B and E) and docetaxel at 100 nM (C and F). Images were digitally scanned ×40 (AlexaSoft X-PRO).

In vivo anti-tumor effect of Y-TR1 in the NOD/SCID mouse xenograft model using the CD26 positive MM cell line JMN. (C) Histological analysis of xenograft tumors of JMN cells. JMN-derived tumors show histopathology of sarcomatoid mesothelioma. (x20). a: Hematoxylin and eosin staining. b: Immunohistochemical staining with anti-human CD26 antibody revealed CD26 expression in tumor cells. c-e: MIB-1 (Ki67) staining showed a decreased number of MIB-1-positive cells in Y-TR1-treated tumors compared to IgG1- or YS110-treated tumors. Scale bar: 10 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31398954), licensed under a CC-BY licence.

T47D cells form functional 3D spheroids in a scaffold-free system. Upper line images show H&E stainings of microtissues grown for 3 days (A) and 12 days (B) after the drop in a GravityTRAP(TM) plate. In addition, sections of 3D microtissues grown for 12 days were incubated with DAPI and an antibody against Ki67 (green) to stain proliferative cells (C) or with an antibody against cleaved Caspase-3 (red) specific to stain apoptotic cells (D) and analyzed by fluorescence microscopy. Nuclei were visualized with DAPI (blue). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26763588), licensed under a CC-BY licence.

HIF-1 alpha binds to the TIM-3 promoter and regulates its expression in primary glia.(a) Cell surface expression of TIM-3 was analysed in BV2 cells under 20% O2 or 1% O2 for 24 h by flow cytometry using PE-conjugated anti-TIM-3 antibody. The results from three independent experiments are presented as a representative histogram and the mean fold change (± s.d.) relative to the normoxic sample. (b) Mouse primary mixed glial cells were incubated under hypoxia or normoxia for 24 h, and the cells were examined by immunocytochemistry using an anti-TIM-3 antibody. (c,d) Mouse primary mixed glial cells and primary neuronal cells were incubated under hypoxia or normoxia for 24 h, and then RT–PCR was used to detect the levels of TIM-3 and actin. Relative transcript levels are shown as the mean fold change (± s.d.) from three independent experiments (NS, not significant, Student–Newman–Keuls test). (e) Primary mixed glial cells were incubated under hypoxia or normoxia for 24 h, and chromatin immunoprecipitation (ChIP) was performed with anti-HIF-1 alpha or control IgG. The results are presented as relative amounts representative of three independent experiments. (f) Primary mixed glial cells were cultured from HIF-1 alpha +f/+f mice, infected with Ad-GFP or Ad-Cre/GFP, transfected with TIM-3-luciferase reporter constructs and incubated under hypoxic or normoxic conditions for 24 h. Relative promoter activity is expressed as the ratio of luciferase activity/ beta -galactosidase activity. (g,h) RT–PCR (g) and western blot analysis (h) were performed under hypoxia or normoxia for 24 h using the indicated primers and antibodies, respectively. The data shown are representative of at least three independent experiments. The graphs show the percent changes in TIM-3 transcript and protein levels in Ad-Cre/GFP- versus Ad-GFP-infected cells under hypoxia. IP, immunoprecipitation. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/25790768), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

A. Co-IP analysis of the interaction between DSP and TRF2 in HEK293 cells and MCF-7 cells. B. Co-IP analysis of the interaction between DSP and TRF2 in DSP-EGFP transfected HEK293 cells or DSP△NLS-EGFP transfected HEK293 cells. C. DSP pull down by telomere targeting CASID with the condition of TRF2 knockdown in HEK293 cells. Histone H3 is the loading control. D.TRF1 and TRF2 pull-down assay by telomere targeting CASID with the condition of DSP knockdown in HEK293 cells. E. Left, Dot blot of telomere DNA repeats in chromatin pull down by DSP with the condition of TRF1 or TRF2 knockdown. The dot-blot was performed at least three times and the represented one is shown. Right, the histogram shows the quantitated results of the dot blots. The triple asterisks (***) indicates a p-value less than 0.01. ns. indicates the insignificance difference. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31595153), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: Rabbit Monoclonal Ki67/MKI67 Antibody (SP6) - Unpurified [NB600-1252] - Image depicting Ki67/MKI67 immunoreactivity in a FFPE section of human tonsil. NB600-1252 was diluted 1-100 and left on tissue sections for 30m at room temperature. Secondary was horse anti rabbit HRP polymer. Image from a verified customer review.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , , , ,

Applications: , , , , ,

Host:

Rabbit Monoclonal

Host: Rabbit Monoclonal

Alt Catalog#

IMG-80336

Alt Catalog# IMG-80336

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Formulations

Formulation	Catalog #	Availability	Price
Unpurified	NB600-1252-0.1ml

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AF3158

KLF4 Antibody [Unconjugated]

(2 Reviews)

(86 Publications)

100 μg

AF3158

D3 mouse embryonic stem cell line was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. KLF4/DNA complexes were immunoprecipitated using 5 μg Goat Anti-Mouse KLF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3158) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The B2R promoter was detected by standard PCR.

KLF4 was detected in immersion fixed D3 mouse embryonic stem cell line using Mouse KLF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3158) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

KLF4 was detected in perfusion fixed frozen sections of mouse colon tissue using Goat Anti-Mouse KLF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3158) at 1 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (brown; Catalog # VC004) and counterstained with hematoxylin (blue). Specific staining was localized to cell nuclei. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Overexpression of beta -catenin maintains B6 ESC self-renewal in the presence of LIF.(A) Immunoblot analysis of phospho-beta -catenin, beta -catenin and beta -actin levels in B6 ES cells cultured in 1000 U/ml LIF condition supplemented without or with 3 µM CH for 4 days. (B) Immunofluorescence analysis of 3 µM CH-treated and non-treated B6 ESC clones stained for beta -catenin and Hoechst for nuclei in the presence of LIF for 4 days. Scale bars represent 100 µm. (C) Quantitative RT-PCR analysis of Axin2, T and Cdx1 transcript levels in 3 µM CH-treated and control B6 ES cells. Error bars represent the SD of three biological replicates. (D) beta -catenin S33A protein stably expressed in ES cells was detected by immunoblotting. (E) Phase contrast microscopy image and alkaline phosphatase staining of control and beta -catenin S33A-transfected B6 ES cells in the presence of 1000 U/ml LIF for 4 days. Scale bars represent 100 µm. (F) Immunofluorescence staining for OCT4, NANOG, and KLF4 in mock and beta -catenin S33A-transfected B6 ES cells in the presence of 1000 U/ml LIF for 4 days. Cell nuclei were stained with Hoechst. Scale bars represent 100 µm. (G) Quantitative RT-PCR analysis of Oct4, Nanog, Rex1, Klf4, Tbx3, and Fgf5 transcript levels in mock and beta -catenin S33A-transfected B6 ES cells in the presence of 1000 U/ml LIF for 4 days. Error bars represent the SD of three biological replicates. (H) beta -catenin S33A delta C, beta -catenin S33A and TCF4 delta N protein were detected by immunoblotting with anti-flag tag antibody from extracts of ES cells transfected with PiggyBac plasmids. Quantitative RT-PCR analysis of Axin2, T and Cdx1 transcript levels in beta -catenin S33A delta C, beta -catenin S33A, TCF4 delta N, and mock-transfected B6 ES cells. Error bars represent the SD of three biological replicates. (I) Alkaline phosphatase staining of mock-transfected, and beta -catenin delta C over-expressing B6 ES cells cultured in LIF for 5 passages, and mock-transfected and TCF4 delta N-transfected B6 ESCs cultured in CHIR99021 plus LIF for 5 passages. Scale bars represent 100 µm. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0035892), licensed under a CC-BY license. Not internally tested by R&D Systems.

LIF maintains B6 ES cell self-renewal in the presence of CHIR99021.(A) Phase contrast image of B6 ES cells in the presence of 1000 U/ml LIF supplemented without or with 3 µM CHIR99021 (CH) for 4 days. Scale bars represent 100 µm. (B) Alkaline phosphatase staining of B6 ES cells in the presence of 1000 U/ml LIF supplemented without or with 3 µM CH for 4 days. Scale bars represent 100 µm. (C) Immunofluorescence staining for OCT4, NANOG, and KLF4 in B6 ES cells treated with 1000 U/ml LIF and 3 µM CH for 4 days. Cell nuclei were stained using Hoechst. Scale bars represent 100 µm. (D) Relative quantification of Oct4, Nanog, Rex1, Klf4, Tbx3, and Fgf5 mRNA in B6 ES cells in 1000 U/ml LIF culture without or with 3 µM CH, by qRT-PCR. Error bars represent the SD of three biological replicates. (E) B6 ES cells,1×104/well, cultured in different concentrations of LIF, ranging from 0 to 10,000 U/ml, in the absence or presence of 3 µM CH for 5 days, were assessed for alkaline phosphatase activity. Scale bars represent 100 µm. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0035892), licensed under a CC-BY license. Not internally tested by R&D Systems.

Overlapping SIM2 occupancy with master transcription factor binding sites.a. Frequency distribution of OCT4, SOX2 and NANOG DNA binding sites in a 40kb window centered to the newly identified SIM2 DNA binding sites. Plots show a significant enrichment for the OSN binding sites at the SIM2 peak localization in SIM2 A6 expressing cells. Pie charts show the proportion of SIM2 DNA binding sites overlapping with the OCT4, SOX2 or NANOG binding sites (in grey) (100bp window). p = Fisher’s exact test p-value; F score: measure of the significance of the association (1 = perfect match). b. Protein co-immunoprecipitation experiments of SIM2-FLAG with endogenous OCT4, SOX2, KLF4 (left panel) and NANOG (right panel). Cellular protein extracts from Sim2 expressing cells (A6) or EB3 cells were immunoprecipitated by using antibodies directed against each of the pluripotency factors (N-terminal and C-terminal part of NANOG) or IgG as a negative control for co-immunoprecipitation. Associated proteins were immunoblotted using an anti-FLAG antibody. Red star shows the SIM2-FLAG protein, blue star the signal given by the recognition of the IgG heavy chains. Ø: Beads only; kDa: kilodaltons; protein lysat: protein lysat was loaded as an input control for the immunoblot. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25955728), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Mu

Species Glossary

Reactivity: Mu

Applications:

, , ,

Applications: , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Biotin	BAF3158

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF3158-SP

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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

Research Areas

Has Sample Size

Promotional Item

Species Glossary

HIF-2 alpha/EPAS1 Antibody - BSA Free

HIF-1 alpha Antibody - BSA Free

LC3B Antibody - BSA Free

SOX2 Antibody [Unconjugated]

Mre11 Antibody

DYKDDDDK Epitope Tag Antibody (L5)

HIF-1 alpha Antibody - BSA Free

FANCD2 Antibody - BSA Free

CHREBP Antibody - BSA Free

TRF-2 Antibody - BSA Free

Ki67/MKI67 Antibody (SP6) - Unpurified

KLF4 Antibody [Unconjugated]