Immunology reagents: Novus Biologicals

NB100-105

HIF-1 alpha Antibody (H1alpha67)

(45 Reviews)

(1064 Publications)

0.1 ml

NB100-105

HIF-1 alpha induction by CoCl2 on Caki-1 cell lysate. WB image submitted by a verified customer review.

<b>Biological Strategies Validation and Genetic Strategies Validation.</b>HIF-1 alpha was detected in immersion fixed DFO treated Hela cells (left) but was not detected in HIF-1 knockout HeLa cells (right) using Mouse Anti-human HIF-1 alpha monoclonal antibody (Catalog # NB100-105) at 25 ug/mL for 3 hours at room temperature. Cells were stained using a NorthernLights (TM) 557-conjugated Donkey Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to nuclei.

<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Naive CD4 T cells from WT, VHL-deficient (Vhl KO), or HIF-1 alpha-deficient (HIF-1 alpha KO) mice were differentiated under IL-22-skewing conditions for a total of 60 h. Some cells remained at normoxia for the duration of the culture (N); others were at normoxia for 35 h and then hypoxia (1% O2) for 24 h (H). At 60 h, nuclear extracts were harvested, and HIF-1 alpha and Lamin B1 levels were analyzed by Western blot. Image from verified customer review.

Staining of HIF-1 alpha in human kidney using NB100-105. Renal tubular epithelium showed moderate membranous, cytoplasmic and nuclear staining, and glomeruli showed faint to moderate nuclear staining.

Analysis using the Alexa Fluor (R) 488 conjugate of NB100-105. Staining of HIF-1 alpha in multiple myeloma cells: H929 cells (0.5 x 10^6) were stained with Alexa Fluor 488(R) conjugated HIF-1 alpha antibody (NB100-105AF488). Image courtesy of Dr. Barbara Muz at Washington University in St. Louis School of Medicine.

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Analysis using the biotin conjugate of NB100-105. Staining of human glioblastoma multiforme.

<b>Biological Strategies Validation. </b>HIF-1 alpha was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with DFOA using 1 ug/mL of mouse anti-HIF-1 alpha monoclonal antibody NB100-105. Cells were stained using a donkey anti-rabbit secondary antibody and counterstained with DAPI (blue).

<b>Biological Strategies Validation. </b>HIF-1-alpha-Antibody-H1alpha67-Western-Blot-NB100-105-img0032.jpg

Analysis using HIF-1 alpha Antibody on human throat cancer tissue. PFA fixed paraffin-embedded sections. Primary antibody dilution 1:100. Image from verified customer review.

<b>Biological Strategies Validation. </b>Analysis of HIF-1 alpha in human hepatocytes from cancer patient using anti-HIF-1 alpha antibody NB100-105. Western blot image submitted by a verified customer review.

<b>Biological Strategies Validation. </b>Analysis using the HRP conjugate of NB100-105. Detection of 50 ug cobalt chloride induced COS-7 nuclear extracts (NB800-PC26) using NB100-105.

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Analysis using the Alexa Fluor(R) 488 conjugate of NB100-105. Staining of HIF-1 alpha in H929 cells using HIF-1 alpha antibody. Flow cytometry image submitted by a verified customer review.

Effects of one- and five-dosage dry needling (1D, 5D) on iNOS, HIF-1 alpha , COX-2, and VEGF expressions in a needling-treated muscle. (a) Representative Western blot images. The quantification of the protein levels for (b) iNOS, (c) HIF-1 alpha , (d) COX-2, and (e) VEGF. Values are expressed as mean ± SD. *Indicates the significant difference (P 50 myofibers from 3 mice/group/time point) & stained for Pax7, DAPI, & EdU (A), HIF2A (B), or HIF1A (C). Scale bars: 20 μm. Arrowheads indicate SCs. (D) Number of Pax7+ SCs per myofiber at various time points. (E) Percentage of EdU+ SCs at various time points. (F) Percentage of HIF2A+ SCs at various time points. (G) Percentage of HIF1A+ SCs at various time points. Data represent the mean ± SEM. Image collected & cropped by CiteAb from the following publication (//www.jci.org/articles/view/96208), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - mTORC2 not mTORC1 regulates HIF‐2 alpha , GOT1 in PDAC under prolonged hypoxia. (A & B) HIF‐2 alpha mRNA & protein levels determined by qRT‐PCR (mRNA) & Western blot (protein) after treatment of Panc‐1 & Capan‐2 cells with mTORC1 inhibitor rapamycin for 48 hrs at 3% or 1% O2. beta ‐Actin was used as loading control. (C & D) Panc‐1 & Capan‐2 cells were treated with mTORC1/mTORC2 inhibitor PP242 & cultured for 48 hrs at 20%, 3% or 1% O2, & HIF‐2 alpha , GOT1 mRNA & protein levels were determined by qRT‐PCR (mRNA) & Western blot (protein). beta ‐Actin was used as loading control. Data are presented as mean ± S.D. from three independent experiments. *P 50 myofibers from n = 3 mice) showing that nmGFP+ QSCs were pimonidazole+. Scale bars: 50 μm & 10 μm (insets). Inset images show that pimonidazole signals were relatively enriched in the cytoplasm of QSCs. Arrowheads indicate a QSC; asterisks indicate a myonucleus. (B) Percentage of pimonidazole+ QSCs. (C) Timeline of in vivo CCI-103F labeling in C57BL/6 mice & representative images of uninjured/resting EDL myofibers (n >50 myofibers from 3 mice) showing that nmGFP+ QSCs were CCI-103F+. Arrowheads indicate a QSC; asterisks indicate a myonucleus. Scale bar: 20 μm. (D) Percentage of CCI-103F+ QSCs. (E & F) Representative images of uninjured/resting EDL myofibers from C57BL/6 mice (n >50 myofibers from 6 mice/group) showing that most QSCs were HIF2A+, but HIF1A–. Scale bars: 10 μm. (G) Percentage of HIF1A+ & HIF2A+ QSCs. Data represent the mean ± SEM. Image collected & cropped by CiteAb from the following publication (//www.jci.org/articles/view/96208), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - FDFT1 inhibits glycolysis through suppressing AKT-mTOR-HIF1 alpha pathway in CRC.aFDFT1 overexpression reduced glucose uptake in CT26 cells. bFDFT1 overexpression decreased lactate production via glycolysis in CT26 cells. c ECAR reduced when FDFT1 overexpressed in CT26 cells. d OCR increased when FDFT1 overexpressed in CT26 cells. eFDFT1 knockdown increased glucose uptake in CT26 cells. fFDFT1 knockdown increased lactate production via glycolysis in CT26 cells. g ECAR increased when FDFT1 knocked down in CT26 cells. h OCR decreased when FDFT1 knocked down in CT26 cells. i, mFDFT1 overexpression inhibited protein & mRNA expression of mTOR-targeted glycolytic enzymes, including GLUT1, HK2, PGK1, GPI, & LDHA, in CT26 cells. j, mFDFT1 knockdown increased protein & mRNA expression of mTOR-targeted glycolytic enzymes, including GLUT1, HK2, PGK1, GPI, & LDHA, in CT26 cells. k, nFDFT1 overexpression decreased protein & mRNA expression of AKT, mTOR, & HIF1 alpha . l, nFDFT1 knockdown increased protein & mRNA expression of AKT, mTOR, & HIF1 alpha . o Photograph of dissected tumors (the first line: normal diet, second line: FMD + glucose, third line: FMD, n = 5). p The tumor volumes measured every 2 days after 13th day. The FMD + glucose group can reverse tumor growth inhibition induced by FMD (n = 5; ns: P = 0.1838; P = 0.0001). q The protein level of FDFT1 & mTOR in dissected tumor samples from normal diet group, FMD group & FMD + glucose group measured by western blotting. r The glucose level in these three groups. Error bars, mean ± SD, data are from three independent experiments. Two-sided t tests. *P < 0.05, **P < 0.01, ***P < 0.001, compared w/ control group (or normal diet group). #P < 0.05, ##P < 0.01. Image collected & cropped by CiteAb from following publication (//pubmed.ncbi.nlm.nih.gov/32313017), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Role of HIF-1 alpha , FOXO3 in hypoxia-induced BNIP3 expression. (A) UCB-hMSCs were incubated with various durations of hypoxia (0–48 h). The protein expressions of HIF-1 alpha & beta -actin were detected by western blot. n = 4. (B) UCB-hMSCs were pretreated with NAC (5 mM) for 30 min prior to hypoxia incubation for 24 h. The protein expressions of HIF-1 alpha , lamin A/C & beta -tubulin in non-nuclear & nuclear fractionized cell samples were assessed by using western blot. n = 3. (C) UCB-hMSCs were immuno-stained with HIF-1 alpha & PI (magnification ×600). Scale bars, 37.5 µm. (D) HIF1A siRNA or NT siRNA was transfected to cells prior to hypoxia treatment for 24 h. The mRNA expression of BNIP3 was analyzed by qPCR. n = 6. (E) The protein expressions of BNIP3 & HIF-1 alpha were detected by western blot. n = 4. (F) NAC (5 mM) was pretreated to UCB-hMSCs prior to hypoxia treatment for 24 h. FOXO3, lamin A/C & beta -tubulin proteins expressions were assessed by western blot. n = 3. (G) FOXO3 siRNA transfected to UCB-hMSCs prior to hypoxia treatment for 24 h. The FOXO3 mRNA expression was measured by qPCR. n = 6. (H) BNIP3, FOXO3 & beta -actin expressions were detected by western blot. n = 3. Western blot data were normalized by beta -actin, & qPCR data were normalized by ACTB mRNA expression level. Lamin A/C & beta -tubulin were used as nuclear & non-nuclear protein controls, respectively. Quantitative data are presented as a mean ± S.E.M. All blots & confocal images are representative. *p < 0.05 versus control, #p < 0.05 versus hypoxia. Image collected & cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S2213231717303804), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - ROS overproduction mediated apoptosis induced by combined treatment of DDP & HIF-1 alpha in PC-3 xenografts & cell culture. (a) ROS monitored by DHE (red) & 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) & culture media (d) were used to detect cellular H2O2 level. (e,f) Western analysis for HIF-1 alpha as well as cleaved caspase-3 & PARP in PC-3 cells following various treatments. Data were presented as mean ± SD of three independent experiments. *p < 0.05 versus control group; #p < 0.05 versus si-HIF-1 alpha or DDP group; $p < 0.05 versus DDP/si-HIF-1 alpha group. The original blots are presented in Supplementary Figure 8. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-07973-4), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - miR-128 downmodulation regulates MAFG increase in hypoxic condition both in vitro & in vivo. C2C12 cells exposed to hypoxia (2% O2) for 4 h, & miR-128 levels (a) determined by RT-qPCR on total RNAs after normalization w/ the small RNU6. Data are expressed as relative to values obtained in untreated control which set equal to 1. Each column in the panels represents the mean ± SD of 3 independent experiments. ∗p < 0.05. Western blotting analysis (b) of HIF-1 alpha , MAFG, & HMOX-1 performed on total extracts from the same cells, as described in Materials & Methods. Tubulin used as a loading control. mRNA levels of HMOX-1 & x-CT genes (c) determined by RT-qPCR on total RNAs, & relative changes calculated by comparing treated cells versus control, after normalization w/ c-ABL. Data reported as relative to values obtained in normoxia, which set equal to 1. Each column in the panel represents the mean ± SD of 3 independent experiments. ∗p < 0.05. Representative laser Doppler analysis (d) of blood flow before & 4 hours after (post 4 h) hindlimb ischemia procedure (n = 3-4 mice/group). Representative Western blot analysis & densitometric analysis (e) of MAFG & HMOX-1 protein levels performed on hindlimb muscle lysates from littermates after sham procedure (sham) or not-ischemic limb (NIL) & ischemic limb (IL) 4 h after femoral artieriectomy. Tubulin protein levels used as loading controls (∗p < 0.05 versus sham; n = 3-4 hindlimb/group). miR-128 levels (f) determined by RT-qPCR on total RNAs after normalization w/ the small RNU6. The values obtained in not-ischemic limb (NIL) set equal to 1. The data are expressed as the mean ± standard error & are representative of 3 independent experiments. ∗p < 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29138682), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Effect of ENMD-1198 on HIF-1 alpha & VEGF expression in HCC cells. To determine effects of ENMD-1198 on nuclear HIF-1 alpha expression, HUH-7 cells were incubated for 16 h ± ENMD-1198 & subsequently exposed to hypoxia (1% O2, 4 h). A) Western blot analysis of whole protein showed that ENMD-1198 effectively blunted hypoxic induction of HIF-1 alpha protein. B) In addition, treatment with ENMD-1198 down-regulated constitutive VEGF-A mRNA levels in HCC cells (HepG2), as measured by real-time PCR (* P < 0.01) (n = 3/group). VEGF-A mRNA expression is normalized to beta -actin. C) ELISA analysis for VEGF in culture supernatants (HepG2). Hypoxia (20 h, 1% O2) markedly increased VEGF protein. ENMD-1198 treatment lowered VEGF secretion under hypoxic conditions (*P < 0.05). D) Western blot analysis for VEGF. Cells were incubated under either non-hypoxic, or hypoxic conditions ± ENMD-1198. Treatment with ENMD-1198 slightly lowered the hypoxic induction of VEGF, as determined by densitometry. Bars: SEM. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/18651980), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Upregulation of HIF-1 alpha in human PCa. (a) HIF-1 alpha protein was detected by western blot in nonmalignant (RWPE-1 & BPH1) & PCa cell lines (PC-3, DU145, LNCaP, & 22RV1) as indicated. (b–d) Total RNA extracted from RWPE-1, BPH1, PC-3, DU145, LNCaP, & 22RV1 cells was subjected to qRT-PCR for HIF-1 alpha (b), VEGF (c) & GLUT4 (d). (e) The HIF-1 alpha promoter-driven reporter (firefly luciferase) & a control vector (Renilla luciferase) were co-transfected into RWPE-1, BPH1, PC-3, DU145, LNCaP, & 22RV1 cells for measurement of luciferase activity. HIF-1 alpha promoter activity was calculated as a ratio of firefly to Renilla activity. (f) Human normal & malignant tissue (Gleason score 9) sections were probed with HIF-1 alpha antibody (scale bars, 100 µm). (g) Protein expression of HIF-1 alpha , VEGF, & GLUT4 were examined with western blot, in PC-3, DU145, & LNCaP cells after various treatments as indicated. Data are expressed as mean ± SD of seven independent experiments. $p < 0.05 versus RWPE-1 or BPH1 cells or normal tissue. *p < 0.05 versus control group. #p < 0.05 versus si-HIF-1 alpha or DDP group. Original blots are shown in Supplementary Figure 5. C: Ctrl; D: DDP; S: si-HIF-1 alpha ; D/S: DDP/si-HIF-1 alpha . Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-07973-4), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Cx43-overexpressing cells regulate the expression of HIF-1 alpha , vascular endothelial growth factor (VEGF), & the proliferation of endothelial cells. (a) The expression of Cx43 & HIF-1 alpha was measured in Cx43-overexpressing cells by Western blot analysis. beta -actin expression served as loading controls for & total protein. Inserted values indicated relative proteins expression in comparison with beta -actin; (b) Cx43-overexpressing cells were cotransfected with pCLNCX-6× HRELuc & pTCYLacZ plasmids. At 6 h post-transfection, their luciferase activities were determined & normalized with beta -gal activity. Data shown were the mean ± SD (n = 4); (c) The conditioned medium of Cx43-overexpressing cells was measured by ELISA. Data shown were the mean ± SD (n = 4); & (d) The conditioned medium of Cx43-overexpressing cells reduced the proliferation of endothelial cells. The HEMC-1 cells treated with conditioned medium of Cx43-overexpressing cells were examined the proliferation activity. Cell viability was assessed by the WST-1 assay. Data shown were the mean ± SD (n = 4). * p < 0.05; *** p < 0.001. Image collected & cropped by CiteAb from the following publication (//www.mdpi.com/1422-0067/16/1/439), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Prolonged hypoxia increases glutamine metabolism in PDAC cells, & HIF‐2 alpha can promote non‐canonical glutamine metabolism in chronic hypoxic conditions. (A) Time course of glutamine consumption at 1%, 3% or 20% O2, each time data point is an average of triplicate experiments. (B) Panc‐1 & Capan‐2 were incubated for 48 hrs at 1%, 3% or 20% O2, GLS1, GOT1 & GOT2 protein were measured by Western blot. beta ‐Actin was used as loading control. (C) Panc‐1 & Capan‐2 were incubated for 48 hrs at 1%, 3% or 20% O2, GLS1, GOT1 & GOT2 mRNA were measured by qRT‐PCR. Data are presented as mean ± S.D. from three independent experiments. (D & F) Si‐HIF‐2 alpha ‐transfected Panc‐1 & Capan‐2 cultured at 1% or 3%O2 for 48 hrs. The level of HIF‐2 alpha & glutamine metabolism enzymes mRNA & protein were determined by qRT‐PCR (mRNA) & Western blot (protein), & beta ‐actin was used as loading control. Data are presented as mean ± S.D. from three independent experiments. *P < 0.05, **P < 0.01. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28544376), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - HIF1 alpha & PFKFB3 are protective against hIAPP mediated beta -cell death. a Representative immunoblotting (left) & quantification (right) of the whole cell extract (WCE) from asynchronous INS 832/13 cells treated with either HIF1 alpha inhibitor KC7F2 (10 μM) or PFKFB3 siRNA (75 nM) for 56 h versus control. Cells were transduced with hIAPP adenovirus (75 MOI) or LacZ adenovirus as control (75 MOI) for last 32 h of culture. DMSO or CTRL siRNA, respectively were added to LacZ or hIAPP to equal DMSO or PFKFB3 siRNA used in experimental groups. GAPDH was used as a loading control. Data are presented as mean ± SEM, n = 4, *p < 0.05, **p < 0.01. b Quantification of DNA content distribution after FACS analysis of INS 832/13 cells treated as described in (a). Data are the mean ± SEM, n = 4, *p < 0.05, **p < 0.01. c The frequency of TUNEL positive INS 832/13 cells treated as described in (a). Data are the mean ± SEM, n = 4 in (a) & (b), & n = 3 independent experiments in (c). Statistical significance was analyzed by one-way ANOVA test with Tukey’s post-test (*p < 0.05, **p < 0.01). See Supplementary Fig. 10 for additional supportive data Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31213603), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Immunohistochemical & Western blot analysis of HIF-1 alpha & VEGF expression in OVA-challenged mice.(A & B) Immunostaining for HIF-1 alpha & VEGF in nasal mucosa following the last challenge. Positive HIF-1 alpha signals are brown & are predominantly nuclear & VEGF is predominantly cytoplasmic (magnification 400×, scale bar = 20 µm). (C & D) Representative Western blot analysis showing HIF-1 alpha & VEGF (with beta -actin as a loading control) expression in the nasal mucosa 24 h after the last challenge. All densitometric analyses are presented as the relative ratio of each molecule to beta -actin & the ratio in negative control mice was set to 100. Values represent means±SEM (n = 6 mice). *Significantly different from negative control, p<0.05; #significantly different from positive control, p<0.05. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0048618), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - DC101, evofosfamide, & HIF-1 alpha shRNA treatment of HT1080 sarcomasA. Western blot of HT1080 cells stably transduced with HIF-1 alpha shRNA (sh.HIF-1 alpha ) or scrambled control shRNA (sh.Scr) & grown in 21% oxygen or 1% oxygen. beta -actin serves as loading control. B. Growth of HT1080 cells stably transduced with sh.HIF-1 alpha or sh.Scr in athymic nude mice. Groups were treated with control IgG 20 mg/kg 3 times per week, DC101 20 mg/kg 3 times per week, and/or evofosfamide (Evo) 50 mg/kg 5 times per week. C. Photos of H&E staining, cleaved caspase 3 IF, cleaved caspase 3 & CD31 IF, CD31 IHC, pimonidazole IHC, & HIF-1 alpha IHC. D. Graphs of microvessel density, pimondizale area, & nuclear HIF-1a expression. Bars represent standard deviation. **p < 0.05 compared to all other groups. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.10212), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - 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 & are expressed relative to control (n = 3). (B,C) Increased Wnt11 mRNA by hypoxia in EMSC preadipocytes & adipocytes (Day 0–12 after differentiation) (B), & C2C12 myoblast & myocyte (Day 0 & 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 & 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) & 4T1 cells (F). (G) EMSCs treated with 0.1 mM DMOG for the indicated times. Wnt11 & Vegf mRNA expression was measured by qPCR & 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, & ERK were used as loading controls, WNT11 normalized to alpha -Tubulin was shown. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - CAPE inhibits MCT-induced HIF-1 alpha expression & PDGF-BB production in rats. (A) Rats were treated with PBS or CAPE (5 or 10 mg/kg) from days 14–28 after MCT treatment 14 days (60 mg/kg). Preparation of lung tissues was analyzed by Western blot to determine the levels of HIF-1 alpha protein. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH)was used as a loading control. (B) Serum levels of PDGF-BB in rat was assayed by an ELISA kit. (C) Representative images of immunohistochemical staining for HIF-1 alpha in lung sections from the rats. Scale bars, 50 μm. (D) Immunofluorescence staining of alpha-smooth muscle actin ( alpha -SMA) & HIF-1 alpha in lung sections from rats. Scale bars, 50 μm. Data in (A,B) are expressed as mean ± SEM of three independent experiments. *** p < 0.01, as compared with the PBS group. ### p < 0.01, as compared with the rats exposed to MCT alone. Results are representative of three rats per experimental group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30909527), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - mTOR expression is inversely correlated with FDFT1 expression.aFDFT1 overexpression inhibited the protein level of mTOR, whereas FDFT1 knockdown increased the protein level of mTOR in CT26 & SW620 cells. bFDFT1 expression & mTOR expression were negatively correlated in CRC patient samples. Scale bar: 200 µm. c The mTOR silencing efficiency of the siRNA in CT26 & SW620 cells was validated by western blotting. d The effect of mTOR silencing on FDFT1 expression level in CT26 was evaluated by western blotting. e The protein level of mTOR when FDFT1 knockdown combined with or without mTOR inhibitor in CT26. f The protein level of pS6k, S6k, pS6, & S6 under the effect of fasting & FDFT1 overexpression in CRC cells. g, h CCK8 proliferation assays showed that the silencing of mTOR decreased the proliferation of CT26 & SW620 cells. k, n The silencing of mTOR reduced glucose uptake & lactate production in CT26 & SW620 cells. i, j, l, o The silencing of mTOR decreased the ECAR & increased the OCR in CT26 & SW620 cells. m The silencing of mTOR decreased the expression of AKT, HIF1 alpha , & proteins encoded by relevant glycolytic genes, such as GLUT1, HK2, LDHA, GPI, PGK1, in CT26 & SW620 cells. p–t Based on TCGA data set analysis, mTOR expression was positively correlated with AKT1, HIF1 alpha , GLUT1, HK2, & LDHA expression. Error bars, mean ± SD, the data are from three independent experiments. Two-sided t tests. *P < 0.05, **P < 0.01, ***P < 0.001, compared with the control group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32313017), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Chromatin Immunoprecipitation: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - HIF-1 alpha -mediated repression of ENT1 expression in Caki-1 cells under hypoxia. Comparison of HIF-1 alpha & HIF-2 alpha protein expression in human matched renal normal-tumor samples (n=11) (A) & 3 RCC cell lines (786-O, 769-P, Caki-1) under normoxia or hypoxia for 6 h (B). (C) Schematic diagram of two hypoxia response elements (HRE) in the promoter of ENT1. (D) CHIP-qPCR analysis of HIF-1 alpha at the promoter of ENT1 in Caki-1 cells, exposed under normoxia or hypoxia for 48 h. Two different HIF-1 alpha antibodies (610958, BD Biosciences; NB100-105, Novus) were used to pull down HIF-1 alpha . The protein expression level of HIF-1 alpha (E) & mRNA expression level of SLC29A1 (F) in Caki-1 under normoxia or hypoxia & HIF-1 alpha knockout-Caki-1 cells. (G) The expression level of ARNT1 (HIF-1 beta ) was detected in Caki-1, transfected with ARNT1 specific siRNAs or NC. Student's t test (two-tailed) was used. (H, I) The expression level of ENT1 in Caki-1, transfected with ARNT1 specific siRNAs or NC, under normoxia or hypoxia. Oneway-ANOVA analysis was used. The results are expressed as mean ± SEM (n=3). ***P<0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32206108), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Upregulation of HIF-1 alpha in the spleen restricts CD8+ T cell expansion.(A-B) Mice were infected with 2x107 amastigotes intravenously & euthanized at various time points after infection. HIF-1 alpha expression in splenocytes was assessed by real-time PCR (A) & immunoblot analysis (B). (C-F) 2x104 OT-I CD8+ T cells were adoptively transferred into recipient mice a day prior to infection with ovalbumin-transgenic (PINK) L. donovani amastigotes. (C) Graphs represent the average number of OT-I CD8+ T cells found in the spleen from Hif1aflox/flox & Hif1a+/-mice at d7 & d14 p.i.. (D) Percentage of OT-I CD8+ T cells at d7 & d14 p.i.. (E) Percentage of gated OT-I CD8+ expressing CD62Llo/int & negative for CD127. (F) Percentage of OT-I CD8+ T cells that were CD44+ CD127+CD62Lhi. All data represent mean ± SEM of one of 3 independent experiments, n = 5. * denotes p<0.05 & ** denotes p<0.01. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.ppat.1004938), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Ccl7 is a HIF1 alpha direct target gene.a qPCR analysis of Ccl7 mRNA expression in Hif1afl/fl & Hif1a delta SMC VSMCs treated with vehicle, CoCl2, normoxia, or hypoxia (2% O2) for 6, 12, & 24 h. *P < 0.05, **P < 0.01, ***P < 0.001, n = 6 (independent experiments) per group. VSMCs isolated from Hif1afl/fl mice & Hif1a delta SMC mice were infected with oxygen-stable HIF1 alpha -expressing lentivirus, & then treated with Ang II for 24 h, bCcl7 mRNA was measured by qPCR & c CCL7 protein was detected by ELISA. d qPCR analysis of Ccl7 mRNA expression in vehicle or Ang II-treated Hif2afl/fl & Hif2a delta SMC VSMCs. e Schematic diagram of the mouse Ccl7 promoter illustrating the HREs in the regulatory region; the upstream regions were numbered in relation to the transcription initiation site. f Luciferase-reporter constructs under the control of the mouse Ccl7 promoter. HEK293T human embryonic kidney cells transiently transfected with the luciferase construct, & cotransfected with empty vector or HIF1a expression plasmids. Standard dual-luciferase assays were performed. EV, empty vector. **P < 0.01, n = 3. g, h ChIP assays of vehicle or Ang II-treated wild-type VSMCs using HIF1 alpha or HIF2 alpha antibodies. Data were normalized to input. *P < 0.05, **P < 0.01, n = 6 per group. i, j ChIP assays of vehicle or Ang II-treated Hif1afl/fl & Hif1a delta SMC VSMCs using HIF1 alpha antibody. Data were normalized to input. *P < 0.05, **P < 0.01, n = 6 per group. Statistical significance was determined by one-way ANOVA test followed by the unpaired t-test Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31320613), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - CAPE inhibits MCT-induced HIF-1 alpha expression & PDGF-BB production in rats. (A) Rats were treated with PBS or CAPE (5 or 10 mg/kg) from days 14–28 after MCT treatment 14 days (60 mg/kg). Preparation of lung tissues was analyzed by Western blot to determine the levels of HIF-1 alpha protein. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH)was used as a loading control. (B) Serum levels of PDGF-BB in rat was assayed by an ELISA kit. (C) Representative images of immunohistochemical staining for HIF-1 alpha in lung sections from the rats. Scale bars, 50 μm. (D) Immunofluorescence staining of alpha-smooth muscle actin ( alpha -SMA) & HIF-1 alpha in lung sections from rats. Scale bars, 50 μm. Data in (A,B) are expressed as mean ± SEM of three independent experiments. *** p < 0.01, as compared with the PBS group. ### p < 0.01, as compared with the rats exposed to MCT alone. Results are representative of three rats per experimental group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30909527), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Role of HIF-1 alpha , FOXO3 in hypoxia-induced BNIP3 expression. (A) UCB-hMSCs were incubated with various durations of hypoxia (0–48 h). The protein expressions of HIF-1 alpha & beta -actin were detected by western blot. n = 4. (B) UCB-hMSCs were pretreated with NAC (5 mM) for 30 min prior to hypoxia incubation for 24 h. The protein expressions of HIF-1 alpha , lamin A/C & beta -tubulin in non-nuclear & nuclear fractionized cell samples were assessed by using western blot. n = 3. (C) UCB-hMSCs were immuno-stained with HIF-1 alpha & PI (magnification ×600). Scale bars, 37.5 µm. (D) HIF1A siRNA or NT siRNA was transfected to cells prior to hypoxia treatment for 24 h. The mRNA expression of BNIP3 was analyzed by qPCR. n = 6. (E) The protein expressions of BNIP3 & HIF-1 alpha were detected by western blot. n = 4. (F) NAC (5 mM) was pretreated to UCB-hMSCs prior to hypoxia treatment for 24 h. FOXO3, lamin A/C & beta -tubulin proteins expressions were assessed by western blot. n = 3. (G) FOXO3 siRNA transfected to UCB-hMSCs prior to hypoxia treatment for 24 h. The FOXO3 mRNA expression was measured by qPCR. n = 6. (H) BNIP3, FOXO3 & beta -actin expressions were detected by western blot. n = 3. Western blot data were normalized by beta -actin, & qPCR data were normalized by ACTB mRNA expression level. Lamin A/C & beta -tubulin were used as nuclear & non-nuclear protein controls, respectively. Quantitative data are presented as a mean ± S.E.M. All blots & confocal images are representative. *p < 0.05 versus control, #p < 0.05 versus hypoxia. Image collected & cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S2213231717303804), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Cx43-knockdown cells regulate the expression of HIF-1 alpha , c-Src, VEGF & the proliferation of endothelial cells. (a) The expression of Cx43, c-Src, & HIF-1 alpha was measured in Cx43-knocknown cells by Western blot analysis. beta -actin expression served as loading controls for & total protein. Inserted values indicated relative protein expression in comparison with beta -actin; (b) Cx43-knockdown cells were cotransfected with pCLNCX-6× HRELuc & pTCYLacZ plasmids. Then, cells were treated with 100 ng/μL PP2 or DMSO for 1 h. At 6 h post-transfection, their luciferase activities were determined & normalized with beta -gal activity. Data shown were the mean ± SD (n = 4); (c) The conditioned medium of Cx43-knockdown cells was measured by ELISA. Data shown were the mean ± SD (n = 4); (d) The conditioned medium of Cx43-knockdown cells reduced the proliferation of endothelial cells. The HEMC-1 cells treated with conditioned medium of Cx43-knockdown cells were examined for proliferation activity. Cell viability was assessed by the WST-1 assay. Data shown were the mean ± SD (n = 4). * p < 0.05; ** p < 0.01; *** p < 0.001. Image collected & cropped by CiteAb from the following publication (//www.mdpi.com/1422-0067/16/1/439), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Involvement of CBP in BNIP3 expression regulated by HIF-1 alpha & FOXO3 under hypoxia. (A) UCB-hMSCs were incubated with hypoxia condition for 24 h. Co-immunoprecipitation of HIF-1 alpha & FOXO3 with IgG & CBP were shown in left panel. IgG was used as a negative control. The total protein expressions of HIF-1 alpha , FOXO3, CBP & beta -actin in lysate were shown in right panel. n = 3. (B) CBP (20 μM) was pretreated to UCB-hMSCs, & cells were incubated with hypoxia for 24 h. The BNIP3 mRNA expression level was analyzed by qPCR. n = 6. (C) BNIP3 & beta -actin protein expressions were analyzed by western blot. Data represent mean ± S.E. n = 4. (D, E) Sample DNA was immuno-precipitated with RNA polymerase, IgG, HIF-1 alpha & FOXO3 specific antibodies. CHIP (top panel) & lysate (bottom panel) samples were amplified with the primers of GAPDH & BNIP3 promoters. Quantitative CHIP data was analyzed by qPCR, & shown in the right panel. n = 4. Western blot data were normalized by beta -actin, & qPCR data were normalized by ACTB mRNA expression level. Quantitative data are presented as a mean ± S.E.M. All blot images are representative. *p < 0.05 versus control, #p < 0.05 versus hypoxia. Image collected & cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S2213231717303804), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Precision quantitative image analysis workflow for the characterization of functional ITH.(a) Representative areas with positive immunohistochemical (IHC) staining for HIF-1 alpha , HIF-2 alpha , phospho-mTOR S2448, phospho-S6RP S235/236 & Ki-67. Only nuclear HIF-1 alpha & HIF-2 alpha expression was included in the analysis. Scale bar, 100 μm. (b) Overview of the quantitative IHC analysis process using a representative image of a whole-slide scanned pT1M1 tumour after phospho-mTOR S2448 staining. The image was overlaid with a virtual grid consisting of 1 mm2 squares that defines the tumour regions to be analysed. Non-cancerous tissue was subtracted manually (dark grey) & a positive pixel count was performed on the corrected area. Positivity per square was expressed as percentage of positive pixels. If a square contained non-tumorous tissue, the positivity was normalized to the corrected square area. Computed values were depicted as a heat map. Coloured squares represent tumour, whereas open squares indicate completely non-tumorous areas. Squares defined as peripheral tumour zone are marked by a black line. Scale bar, 1 cm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27291893), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Regulation of OA development by HIF-1 alpha . Plain radiographs of the entire bodies at 8 weeks of age (A), & knee joints 8 weeks after OA surgery (B) of Cntl & cKO littermates. Scale bars, 10 mm & 1 mm, respectively. (C) Safranin O & TUNEL staining’s of Cntl & KO knee joints at 8 weeks after OA surgery (n = 9 & 11, respectively). Tamoxifen induction was performed at 7 weeks. Inset boxes in top images indicate the regions of high magnification Safranin O & TUNEL images. Inset boxes in middle images indicate the regions of immunofluorescence in (G). White dotted lines in TUNEL images indicate the cartilage area. Scale bars, 400 µm & 100 µm, respectively. (D) OARSI (Osteoarthritis Research Society International) score of OA development. Lines show the mean ± SD. *P < 0.05. (E) Rate of TUNEL-positive cells in the cartilage area. Bars show the mean ± SD. *P < 0.05. (F) mRNA levels of Hif1a in articular cartilage tissues of Cntl & KO. Bars show the mean ± SD. *P < 0.05. (G) Immunofluorescence of Hif1a, Mmp13, Hif2a, I kappa B alpha , Ser 32 & 36 dual phosphorylated I kappa B alpha , Rela, & C1qtnf3. Scale bars, 20 µm. The percentage of positive cells in the immunofluorescence is shown below. *P < 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32214220), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - A beta facilitates HIF1 alpha synthesis & autophagy inhibition via mTOR activation. (A) SK-N-MC cells exposed to A beta (5 μM) for 0–48 h. HIF1 alpha & beta -actin expression analyzed by WB. n = 3. (B) Cells pretreated w/ NAC (1 mM) for 30 min prior to A beta treatment for 24 h. HIF1 alpha & beta -actin expression analyzed by WB. n = 3. (C,E) Cells incubated w/ rapamycin (10 nM) for 30 min prior to A beta treatment for 24 h. Phosphorylation of 4EBP1 (Thr 37/46) & 4EBP1, phosphorylation of p70S6K1 (Thr 389), HIF1 alpha & beta -actin analyzed by WB. n = 6. (D) Protein samples immunoprecipitated by eukaryotic translation initiation factor 4E (eIF4E) antibody-conjugated protein A/G agarose beads. Samples blotted w/ 4EBP1 & eIF4E-specific antibodies. n = 3. (F) Cells exposed to PF4708671 (10 μM) for 30 min prior to A beta treatment for 24 h. HIF1 alpha & beta -actin expression detected by WB. n = 6. (G) Cells exposed to cycloheximide (4 μM) for 30 min prior to A beta treatment for 24 h. HIF1 alpha & beta -actin expressions detected by WB. n = 6. (H) Cells pretreated w/ rapamycin (10 nM) for 30 min, incubated w/ A beta for 24 h & analyzed by WB w/ LC3, p62 & beta -actin specific antibodies. n = 3–6. (I) LC3 puncta visualized by confocal microscopy. Presented results merged images. Green & red fluorescents indicate LC3 & PI respectively. Scale bars, 50 μm (magnification × 600). (J) Cells pretreated w/ trehalose (10 μM) for 30 min prior to A beta treatment for 24 h. Cytotoxicity measured by MTT assay at an absorbance of 545 nm using a microplate reader. Data present the mean ± SE. n = 6. (K) Cell viability measured by trypan blue exclusion assay. Data presented as a mean ± SE. n = 6. Each blot image presented as representative image. *p < 0.05 vs. control, #p < 0.05 vs. A beta treatment. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28790888), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - TSA reduces HIF-1 alpha & VEGF expression & up-regulates the expression of PEDF.Real-time PCR (A) & Western blot assays (B) were performed on RPE cells treated with 0–0.5 μM TSA & 150 μM CoCl2. (A) Cells were treated with 0–0.5 μM TSA for 14 h & then co-treated with 150 μM CoCl2 for 6 h for the analysis of gene expression by real-time PCR. Changes in HIF-1 alpha mRNA levels were not statistically significant. CoCl2 causes a fourfold enhancement of VEGF mRNA expression; but at 0.5 μM TSA, the mRNA level of VEGF reduces to less than half of that in cells treated with CoCl2 only. TSA induces a statistically significant increase in the mRNA level of PEDF. (B) Cells were treated with 0–0.5 μM TSA for 18 h & then co-treated with 150 μM CoCl2 for 6 h for Western blot analysis. TSA reduces the CoCl2-induced HIF-1 alpha & VEGF protein levels by 4.3-fold & 5.7-fold, respectively, & up-regulates PEDF protein level by threefold. (C) Densitometry data for Western blot of HIF-1 alpha , VEGF & PEDF. (*: t test p<0.05; **: t test p<0.01). Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0120587), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Role of HIF-1 alpha , FOXO3 in hypoxia-induced BNIP3 expression. (A) UCB-hMSCs were incubated with various durations of hypoxia (0–48 h). The protein expressions of HIF-1 alpha & beta -actin were detected by western blot. n = 4. (B) UCB-hMSCs were pretreated with NAC (5 mM) for 30 min prior to hypoxia incubation for 24 h. The protein expressions of HIF-1 alpha , lamin A/C & beta -tubulin in non-nuclear & nuclear fractionized cell samples were assessed by using western blot. n = 3. (C) UCB-hMSCs were immuno-stained with HIF-1 alpha & PI (magnification ×600). Scale bars, 37.5 µm. (D) HIF1A siRNA or NT siRNA was transfected to cells prior to hypoxia treatment for 24 h. The mRNA expression of BNIP3 was analyzed by qPCR. n = 6. (E) The protein expressions of BNIP3 & HIF-1 alpha were detected by western blot. n = 4. (F) NAC (5 mM) was pretreated to UCB-hMSCs prior to hypoxia treatment for 24 h. FOXO3, lamin A/C & beta -tubulin proteins expressions were assessed by western blot. n = 3. (G) FOXO3 siRNA transfected to UCB-hMSCs prior to hypoxia treatment for 24 h. The FOXO3 mRNA expression was measured by qPCR. n = 6. (H) BNIP3, FOXO3 & beta -actin expressions were detected by western blot. n = 3. Western blot data were normalized by beta -actin, & qPCR data were normalized by ACTB mRNA expression level. Lamin A/C & beta -tubulin were used as nuclear & non-nuclear protein controls, respectively. Quantitative data are presented as a mean ± S.E.M. All blots & confocal images are representative. *p < 0.05 versus control, #p < 0.05 versus hypoxia. Image collected & cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S2213231717303804), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - HIF1 alpha protein is suppressed by ED71 but not by 1,25(OH)2D3.(A) Western analysis of Raw264.7 cells cultured in hypoxic conditions with or without 10−7 M of ED71 or 1,25(OH)2D3 (1,25D). (B) Hif1 alpha mRNA levels in Raw264.7 cells cultured in hypoxic conditions were analyzed by realtime PCR in the presence or absence of 10−7 M ED71 or 1,25(OH)2D3. Data represent mean Hif1 alpha expression relative to that of Actb ± SD (n = 5). (C) Levels of VDR transcripts in Raw264.7 cells transfected with shRNA targeting the VDR (shVDR) or control shRNA (Control) were determined by realtime PCR. Data represent mean VDR expression relative to that of Actb ± SD (n = 5). (D) Western analysis of control (shControl) or VDR-suppressed (shVDR#1 or shVDR#2) Raw264.7 transformants cultured in hypoxic conditions with ED71 or 1,25(OH)2D3 (1,25D), both at 10−7 M. (E) M-CSF-dependent Ctsk Cre/Hifflox/flox cells were cultured in normoxic conditions to suppress HIF1 alpha in the presence of M-CSF (50 ng/ml) plus RANKL (25 ng/ml) with either ED71 or 1,25(OH)2D3 (1,25D) both at 10−7M for 4 days. Expression of Ctsk & NFATc1 was then assessed by realtime PCR. Data represent mean Ctsk or NFATc1 expression relative to that of Actb ± SD (n = 5). *P<0.05; **P<0.01; ***P<0.001. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0111845), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Lack of HIF-P4H-1 increases the amount of p53. (a) Western blot analysis of p53 in wt & Hif-p4h-1−/− MEFs cultured in 21% or 1% O2 for 24 h. (b,c) Western blot analysis of p53 in Hif-p4h-1 & scrambled (Scrm) siRNA transfected HEK293 cells (b) & wt MEFs treated with 50 µM FG4497 for 24 h (c). (d) qPCR analysis of p53 mRNA in wt & Hif-p4h-1−/− MEFs cultured in 21% or 1% O2 for 24 h. (e) Analysis of p53 protein turnover rate. Hif-p4h-1−/− & wt MEFs were treated with 200 μg/ml of cycloheximide for the indicated time points & whole cell lysates were blotted for p53. (f) Western blot analysis of MDM2 in wt & Hif-p4h-1−/− MEFs cultured in 21% or 1% O2 for 24 h. (g) Western blot analysis of p53 & MDM2 in wt MEFs treated with 10 μM nutlin-3a for 24 h with or without overexpression of human HIF-P4H-1 (OE). (h,i) Western blot analysis of HIF1 alpha in wt & Hif-p4h-1−/− MEFs cultured in 21% or 1% O2 for 24 h (h) & in scrambled & Hif-p4h-1 siRNA transfected HEK293 cells (i). (j) Western blot analysis of ubiquitination of p53 in Hif-p4h-1 & scrambled siRNA transfected HEK293 cells. The cells were transfected with HA-ubiquitin & endogenous p53 was immunoprecipitated followed by Western blotting with anti-HA & anti-P53 antibodies. (k) Western blot analysis of p53 in Hif-p4h-1−/− & wt MEFs were treated with or without 10 μM MG132 for 5 h. Data are presented as representative Western blots & as mean ± s.d., n = at least 3 individual MEF isolates or experiments. *P < 0.05, **P < 0.01 & ***P < 0.001, two-tailed Student’s t-test. Unprocessed original scans of blots are shown in Supplementary Fig. 5. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-17376-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - NDRG1 was regulated by HIF‐1/2 alpha . (A‐E) The protein expression of the shown genes was detected by Western blot, respectively; beta ‐actin was used as the loading control. (A) RCC4 & 786‐O cells (B) were infected with VHL vector (VHL) or empty vector (EV). (C) Caki‐1 cells were transfected with VHL shRNAs (shVHL) or non‐specific control (NC). (D) RCC4 cells were infected with HIF‐1 alpha siRNAs (siHIF‐1 alpha ) or non‐specific control (NC). (E) 786‐O cells were transfected with HIF‐2 alpha shRNAs (shHIF‐2 alpha ) or non‐specific control (NC). (F) Representative images of HIF‐1 alpha , HIF‐2 alpha & NDRG1 protein expression from the IHC staining of human clear cell renal cell cancer specimens. Patient I: The protein levels of HIF‐1 alpha , HIF‐2 alpha & NDRG1 were low. Patient II: The protein levels of HIF‐1 alpha , HIF‐2 alpha & NDRG1 were medium. Patient III: The protein levels of HIF‐1 alpha , HIF‐2 alpha & NDRG1 were high (scale bar, 50 μm). (G & H) The protein level of NDRG1 is significantly correlated with HIF‐1 alpha (G) & HIF‐2 alpha (H). The protein levels were evaluated according to IHC scores, representing very low (score 0‐2), low (score 3‐4), high (score 5‐8) & strong (score 9‐12). The subjects were divided into four groups according to the IHC scores of HIFs in the tumours. (I) The protein level of NDRG1 is significantly correlated with the average of HIF‐1 alpha & HIF‐2 alpha in ccRCC tissues. The protein levels are evaluated according to IHC scores, representing very low (score < 3), low (score < 5), high (score < 8) & strong (score 8‐12). The subjects were divided into four groups according to the IHC scores of the average of HIF‐1 alpha & HIF‐2 alpha . Significance p‐values & r values according to Spearman's rank correlation Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32537867), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Bv, Ca, Fe, Ma, Pm, Pm, Rb, Sh, Xp

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Ca, Fe, Ma, Pm, Pm, Rb, Sh, Xp

Applications:

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

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

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-105AF350
Alexa Fluor 405	NB100-105AF405
Alexa Fluor 488	NB100-105AF488
Alexa Fluor 532	NB100-105AF532
Alexa Fluor 594	NB100-105AF594
Alexa Fluor 647	NB100-105AF647
Alexa Fluor 700	NB100-105AF700
Alexa Fluor 750	NB100-105AF750
Allophycocyanin	NB100-105APC
Biotin	NB100-105B
CoraFluor 1	NB100-105CL1
DyLight 350	NB100-105UV
DyLight 405	NB100-105V
DyLight 488	NB100-105G
DyLight 550	NB100-105R
DyLight 594	NB100-105DL594
DyLight 650	NB100-105C
DyLight 680	NB100-105FR
DyLight 755	NB100-105IR
FITC	NB100-105F
HRP	NB100-105H
Janelia Fluor 525	NB100-105JF525
Janelia Fluor 549	NB100-105JF549
Janelia Fluor 646	NB100-105JF646
Janelia Fluor 669	NB100-105JF669
PE	NB100-105PE
PerCP	NB100-105PCP
mFluor Violet 450 SE	NB100-105MFV450
mFluor Violet 500 SE	NB100-105MFV500
mFluor Violet 610 SE	NB100-105MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Azide and BSA Free	NBP2-80760

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NB100-479

HIF-1 alpha Antibody - BSA Free

(28 Reviews)

(546 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 (*p N) localization in three microscopic viewing areas were counted as shown in the lower panel. The total counted cell numbers ranged from 35 to 64. The experiments were repeated more than twice, except for the cell localization count, & the results were reproduced. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28775317), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-479] - Hypoxia signaling & cytotoxicity to CdCl2. (a) Wild-type (WT) cells were left untreated (Ctrl) or exposed to 150 μM CoCl2 (Co2+), 1, 5 or 10 μM CdCl2 (Cd2+) or 150 μM CoCl2 & 10 μM CdCl2 for 24 hours. HIF1 alpha −/− cell extract treated with 150 μM CoCl2 was used as a negative control. Nuclear protein was extracted & separated by SDS-PAGE, transferred to nitrocellulose membrane & probed with a HIF1 alpha (upper panel) or beta -actin (lower panel) specific antibody. The bands observed in the cadmium only WT cells are nonspecific as they are also observed in the HIF1 alpha −/− cells. (b) BNip3 mRNA expression levels were analyzed by qRT-PCR in wild type (WT, white bars) & HIF1 alpha −/− cells (black bars). Cells were left untreated (0), or exposed to 5 μM CdCl2 (Cd2+) or 150 μM CoCl2 (Co2+) for 24 hours. Each value was normalized to the control level in the corresponding cell line. *P 0.05; *p 60 recipients/genotype). (C) When 500 WT & KO cells were implanted into the mammary fat pad, the median time until 50% of recipients developed tumors > 500 mm3 was 64 days for WT mice & 127 days for KO mice (n = 14 recipients/genotype, P < 0.001, logrank test). (D) Western blot for HIF-1 alpha in three independent tumors (500 to 750 mm3) per genotype. WT & KO cells that were cultured for 6 hours under hypoxic conditions served as positive & negative controls, respectively. CRM, cross-reactive material. MTEC, mammary tumor epithelial cell. (E) Mean fold change ± SEM in expression of HIF-1 targets in KO tumors as determined by qRT-PCR (n = 5 tumors/genotype). Decreased gene expression in KO tumors is presented as a negative fold-change relative to WT tumors. (F) An increase in Ki67+ cells in KO tumors is balanced by an increase in caspase 3-positive cells (n = 5 tumors/genotype, *P < 0.05, Student's t-test). Representative immunostaining images are shown in Additional file 2 Figure S4. Image collected & cropped by CiteAb from the following publication (//breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3087), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-479] - Effect of HIF-1 alpha silencing on alpha -MSH-induced apoptosis in B16-F10 melanoma cells during hypoxia.a Cells were transfected with shLacZ or shHIF-1 alpha plasmids for 48 h before harvest. HIF-1 alpha shRNA reduced the basal HIF-1 alpha mRNA & protein expression levels. b, c Relative mRNA expression levels were analyzed by real-time PCR. Data are expressed as fold change compared with control (means ± SD of triplicate experiments). d Cell lysates were analyzed by immunoblot using the indicated antibodies. beta -Actin was used as an internal control for loading & transfer. e The population of apoptotic cells was analyzed by flow cytometry & qualified as mean ± SD from triplicate experiments. *p < 0.05, **p < 0.01. Knockdown of HIF-1 alpha diminished alpha -MSH-induced apoptosis in B16-F10 melanoma cells during hypoxia Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30062060), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody - BSA Free [NB100-479] - Hif1 alpha expression in neonatal & 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) & 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 & intestinal tissue was used as a negative control. Loading control ( beta -Actin) is shown below. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27148974), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-479] - Expression of hypoxia-inducible factor (HIF) proteins after bilateral renal ischemia or sham ischemia. Immunoblots for HIF-1 alpha (A–C) & HIF-2 alpha (D–F) of tissue extracts from the cortex & outer & inner medulla of the left kidneys of rats 24 h & 5 days following recovery from either sham ischemia (○) or bilateral renal ischemia (●); n = 6 per group. G: typical image of the gel following electrophoresis. H: typical image of the nitrocellulose membrane following transfer. Values are expressed as medians (25th percentile, 75th percentile). Paired comparisons were performed using the Mann-Whitney U-test. Because paired comparisons were made at two time points, P values were conservatively adjusted using the Dunn-Sidak method with k = 2. PTr, PT, & PTr*T are the outcomes of two-way analysis of variance on ranking with the factors treatment (Tr) & time (T). AU, arbitrary unit; I1, 24 h after ischemia; I5, 5 days after ischemia; S1, 24 h after sham ischemia; S5, 5 days after sham ischemia. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30110566), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-479] - Hypoxia reduces inflammatory signaling pathways & NLRP3 expression & induces autophagy in IECs. a HT-29 cells were subjected to normoxia & 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 & the total amount of LC3 (LC3-I plus LC3-II) relative to beta -actin. Results are representative of two independent experiments. b, c & d HT-29 cells were subjected to normoxia & 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 & absence of 10 µg/ml LPS & 20 µM of MG132. Results are representative of two independent experiments. f & g Putative binding sites for HIF-1 alpha & NF-kappa B were found in the p62 f & 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 & 24 h. ChIP analysis was performed using antibodies against HIF-1 alpha & NF-kappa B for immunoprecipitation. PCR was performed using the promoter-specific primers for the p62 f & NLRP3 g promoter binding sites of HIF-1 alpha & 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 & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28740109), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-479] - Transient hyperoxia does not sensitize normal human astrocytes to radiation.Results are shown for the colony forming assays for all previously assayed cell lines & a normal human astrocyte cell line (Astro). Cells were continuously maintained under normoxic conditions (NOx) or exposed to 25 min of hyperoxia (50% O2) & then returned to normoxic conditions for 25 min (HyperOx) before being treated with a 5 Gy dose of radiation. To allow for ease of comparisons among cell types, raw values are expressed as a percentage of the corresponding cells type’s negative (non-irradiated) control & the means & SEMs are plotted. Each result represents three independent samples, plated in triplicate (*p<0.05, Student’s t-test). Also shown are Western blots of nuclear HIF-1 alpha at the time of irradiation for each cell line. Corresponding Western blots of lamin A/C are shown as a loading control. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25350400), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-479] - YC-1 attenuates pro-inflammatory differentiation of macrophages. (a,b) qRT-PCR analyses, revealing mRNA expression of the pro-inflammatory M1-differentiation markers NOS2 (a) & IL-6 (b) in murine J774A.1 macrophages under normoxia (left bars) & hypoxia (right bars). LPS exposure was carried out to induce M1-polarization in presence of YC-1 or vehicle control (DMSO; **P ≤ 0.01, n = 9). (c,d) Western blots of nuclear extracts, revealing protein levels of HIF-1 alpha (c) & HIF-2 alpha (d) in J774.A1 macrophages treated with vehicle (DMSO) or YC-1 under normoxic (left) or hypoxic (right) culture conditions. Simultaneous LPS-treatment was performed to induce pro-inflammatory activation. Note LPS-induced stabilization of HIF-1 alpha - (c), but not HIF-2 alpha -levels (d), & reversal of LPS-induced HIF-1 alpha -stabilization by YC-1 (c). Histone H3 was used as a loading control. (e,f) qRT-PCR analyses, revealing mRNA expression of the pro-inflammatory M1-differentiation markers NOS2 (e) & IL-6 (f) in BMDMs under normoxia (left bars) & hypoxia (right bars). LPS exposure was carried out to induce M1-polarization in presence of YC-1 or vehicle control (DMSO; ***P ≤ 0.001, **P ≤ 0.01, n = 6). Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-13638-z), 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
CoraFluor 1	NB100-479CL1
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 525	NB100-479JF525
Janelia Fluor 549	NB100-479JF549
Janelia Fluor 646	NB100-479JF646
Janelia Fluor 669	NB100-479JF669
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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NB100-449

HIF-1 alpha Antibody

(28 Reviews)

(324 Publications)

0.1 ml

NB100-449

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

HIF-1 alpha /2 alpha expression in myeloid-specific KO mice targeting the HIF pathway. (A) Images of the colon of wild-type (WT), myeloid-specific Hif-1a KO (hMRP8 Hif-1a KO) or von Hippel Lindau (Vhl) KO (hMRP8 Vhl KO) mice, immunostained for MRP8 (green) & the DNA-binding regions of Hif-1a mRNA (red). Mice were fed with 5% DSS for 4 days prior to immunostaining analyses. Note that there were no MRP8-positive cells that were positive for Hif-1a mRNA in hMRP8 Hif-1a KO (middle column) mice, but we observed many cells that were double positive for MRP8 & Hif-1a mRNA in hMRP8 Vhl KO mice (right column). (B) Images of the colon of hMRP8 Vhl KO mice fed with 5% DSS as in A, immunostained for MRP8 (green) & HIF-1 alpha (red, upper row) or HIF-2 alpha (red, bottom row). DAPI-stained nuclei are shown in blue. White boxes in A & B indicate the regions magnified in the lower or right images, respectively. Yellow arrowheads in A & B indicate cells positive for both markers. Scale bars: 100 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29967068), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody [NB100-449] - Analysis of microvascular density & HIF-1 alpha activity. Microvascular density of pulmonary tumors was significantly higher in hyperplastic (B) & tumoral lesions (D) of NNK/NTHi treated mice compared to the NNK treated mice (A, & 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), & high expression in perivascular-peribronchiolar lymphocytes (H). In contrast, low, homogenous expression of HIF-1 alpha was detected in the tumors (E) & perivascular-peribronchiolar lymphocytes (G) of NNK treated mice (40X, scale bar = 25 μm). Image collected & cropped by CiteAb from the following publication (//molecular-cancer.biomedcentral.com/articles/10.1186/1476-4598-11-4), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody [NB100-449] - Chronic ethanol feeding enhances tumorigenesis markers during AOM/DSS-induced tumorigenesis. AOM/DSS-induced colonic tumorigenesis was induced with (EF) or without (PF) 4 % ethanol feeding as described in Methods section. Cryosections of distal colon were stained for VEGF (a) pSmad (b), or HIF1 alpha (c), & co-stained for F-actin & nucleus. Images presented are representative of n of 5 for each group Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26951793), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody [NB100-449] - Treatment with HBOT or PFD decreased HIF-1 alpha expression in the AD lesions.IHC for HIF-1 alpha (A). Scale bar is 100 µm. HIF-1 alpha + area was measured in five fields chosen at random by using image analysis software (B). RT-PCR for HIF-1 alpha (C). NT, not treated. Data are expressed as the mean ± SD (n = 12). *P<0.05, compared with the control group; ¶P<0.05, compared with the AD-NT group; §, P<0.05, compared with the AD-Steroid group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25275529), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody [NB100-449] - Elevated PIK3/AKT/HIF-1 alpha pathways in KL SqCC tumours.(a,b) Analysis of TCGA gene expression (normalized TPM) & PIK3CA (a) & PTEN (b) genomic copy number alteration profiles. Each dot represents one SqCC patient (n=501). Boxes represent the interquartile range & whiskers are drawn to the minimum & maximum. Kruskal–Wallis non-parametric ANOVA, ****P<0.0001, ***P<0.001, **P<0.01. (c) IHC analysis (top) & quantification (bottom) of p63, p-AKT, p-S6, p-4EBP1, HIF-1 alpha & GLUT1 in KL tumours (n=6 each group). Two-tailed t-test, ****P<0.0001, ***P<0.001, **P<0.01. Scale bar, 50 μm. (d) Immunoblot analysis of HIF-1 alpha & GLUT1 in control shGFP & shHIF-1 alpha knockdown SqCC cell lines, HCC95 & HCC1588. All error bars represent the mean±s.e.m. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28548087), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody [NB100-449] - Immunolocalization of HIF1A in limbs.Immunoreactivity was detected in control limbs in the apical ectodermal ridge & interdigital regions (I.R.). The mesenchymal condensations/developing cartilaginous anlagen (Digits) were not immunoreactive. 4-OOHCPA treatment resulted in a concentration- & time-dependent increase in HIF1A immunoreactivity in this area in the 3 & 6 h exposure groups; staining was diminished by 24 h. Four separate replicates were done. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0051937), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Effects of cell stress on VEGF-A & HIF1 alpha expression in Müller cells & Y79 photoreceptors. (A) VEGF-A was measured by ELISA using conditioned media collected from MIO-M1 & Y79 cells, n = 4–5/group in Müller cells & n = 8/group in Y79 photoreceptors, ** p < 0.01; (B) Western blots for HIF1 alpha using cellular proteins, n = 4/group, ** p < 0.01. For MIO-M1 Müller cells: Grey bars: 5 mM glucose. Black bars: 25 mM glucose. For Y79 photoreceptors: Grey bars: 11 mM glucose. Black bars: 25 mM glucose. NS, not significant; LG, low glucose; HG, high glucose. Image collected & cropped by CiteAb from the following publication (//www.mdpi.com/1422-0067/18/3/533), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Phenylhydrazine administration results in hypoxia in retinal vessels & in HIF-1 alpha & HIF-2 alpha stabilization in peripheral blood leukocytes.(A) 3-dimensional reconstructed imaging of the superficial vascular plexus of flat mounted retinae of mice (steady state, treated with PHZ & treated with PHZ & peak EIU induction) stained with DAPI, hypoxyprobe & Isolectin B4. (B) Representative analysis of HIFs by western blot: 50ug of protein from total cell lysate of blood leukocytes isolated from mice treated with PHZ compared to untreated; (C) relative densitometry quantification of HIF western blots from PBMC showing mean, n = 3, error bar showing SD, unpaired t test, ***P < 0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28112274), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Frozen: HIF-1 alpha Antibody [NB100-449] - Ethanol elevates tumorigenic markers & nuclear localization of HIF1 alpha in colonic crypts during AOM/DSS-induced tumorigenesis: AOM/DSS-induced colonic tumorigenesis was induced with or without 4 % ethanol feeding as described in Methods section. a & b Colonic mucosal extracts were immunoblotted for pSmad & VEGF (a), & the band density evaluated using Image J software (b). Values are mean ± SE (n = 3). Asterisks indicate the values that are significantly different (p < 0.05) from corresponding values for AOM/DSS group. c Cryosections of colon were stained for HIF1 alpha (red) & nucleus (blue). Sections of the images from Fig. 2c are enlarged to show the detailed localization of HIF1 alpha in the nucleus. While arrows indicate nuclear co-localization of HIF1 alpha Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26951793), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Dectin-1-induced NADPH oxidase-derived ROS is required for augmented IL-1 beta during TSLPR inhibition. (A) Human mDC derived from healthy donors (HD) or CGD patients were stimulated with SC glucan for 8 h (n = 2 representative donors presented, three separate experiments performed). Pro-IL-1 beta , IL-1 beta , HIF-1 alpha , phospho-AMPK, AMPK & beta -actin were measured by immunoblot. (B–E) Densitometry of cumulative data was performed using Image Studio Lite software with pro-IL-1 beta , IL-1 beta & HIF-1 alpha normalized to beta -actin & phospho-AMPK normalized to total AMPK. Data is reported as percentage of maximal signal observed within each donor (n = 3 independent donors, presented as pooled data). (F–H) Human mDC derived from HD or CGD patients were stimulated with SC glucan, CA glucan or heat killed C. albicans hyphae with anti-TSLPR or IgG isotype control antibodies for 24 h (n = 3 independent donors, presented as pooled data). IL-1 beta was measured in 24-h cell culture supernatants by ELISA. Cumulative data displayed as mean +SEM. Statistical analysis calculated using one-way ANOVA with Bonferroni post-tests (***p = 0.001,*p = 0.05). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31139177), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Reactive oxygen species production & expression of stabilized HIF1 alpha & AMPK subunit alpha 2 & in H1299 & H1299r cells(A) H1299, H1299r, P31 & P31r cell were seeded at a density of 250,000 cells/well & left for 24 h. The cells were then stained with 10 μmol/L dichloro-dihydro-fluorescein diacetate (DCFH-DA). Fluorescence was analysed for 10,000 events. Whole cell lysates were prepared from confluent cultures of H1299, H1299r cells. Proteins (90 μg) were resolved in 10% SDS-PAGE gels & transferred to a PVDF membrane. (B) Representative blot of HIF1 alpha protein expression. (C) Representative blot of AMPK subunit alpha 2 protein expression from four independent experiments. (D) Bar chart of AMPK subunit alpha 2 mean for four separate measurements. Statistical analyses was carried out using the student t-test. ** = p<0.01, * = p<0.05. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.21885), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Pyruvate kinase isoform M2 (PKM2) & hypoxia-inducible factor 1 alpha (Hif-1 alpha ) bind directly to the promoter region of PDL1 in primary murine BMDM cells. BMDM cells were treated with TEPP-46 at 50 µM for 1 h prior to stimulation with LPS (100 ng/ml, 24 h) (A). Binding of PKM2 (left) or Hif-1 alpha (right) to HRE1 or HRE4 of the PD-L1 promoter was detected by incubating cell lysates with biotinylated oligonucleotides spanning the relevant HRE promoter region. Protein–oligonucleotide complexes were isolated using streptavidin agarose beads, & proteins were detected by western blotting. Representative of n = 3. Chromatin immunoprecipitation (ChIP)-PCR (B) using PKM2 & HIF-1 alpha antibodies & primers specific for three promoter regions of PD-L1 (HRE1, 2–3, & 4) & a known Hif-1 alpha binding region of vascular endothelial growth factor as a positive control showing binding of Hif-1 alpha (left) & PKM2 (right) to the PD-L1 promoter in LPS-treated BMDMs (100 ng/ml, 24 h). (C) Sequential ChIP assays measuring simultaneous endogenous binding of PKM2 & Hif-1 alpha to chromatin in response to LPS (100 ng/ml, 24 h) ±TEPP-46 (pretreatment using 50 µM, 60 min). ChIP data are calculated as percent of input, error bar represents mean ± SEM, & statistics are performed as two-tailed unpaired t-test *P < 0.05, **P < 0.01, & ***P < 0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29081778), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Changes in Hepatic Oxygen Saturation & Hypoxia-related Protein Expression. (A) In the representative case, the hepatic oxygen saturation in the experimental group was lower than that in the control group at four, six, & eight weeks (51.6 vs. 54.5%, 48.4 vs. 53.9%, 32.3 vs. 43.2%, & 31.2 vs. 42.9%). (B) There was significant decrease of oxygen saturation of liver parenchyma in the experimental group compared to those of control group (45.2 vs. 51.9%, 43.0 vs. 49.0%, & 31.3 vs. 49.7%) at the four, six, & eight weeks, respectively (P < 0.001). (C) In the Western blot analysis, the experimental group was relatively higher expression of hypoxia-inducible factor-1 alpha (HIF-1 alpha ) & vascular endothelial growth factor (VEGF) protein than the control group at eight weeks. The full-length blots with these antibodies were presented in supplementary Figure S1. (D,E) The experimental group showed significantly higher expression of HIF-1 alpha & VEGF protein than the control group at eight-week (1.47 ± 0.48 vs. 0.19 ± 0.08 & 0.96 ± 0.23 vs. 0.33 ± 0.06, P = 0.019 & P < 0.018). *P < 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29079853), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - BCAT1 expression & inhibition in human macrophages.(a) RNA-sequencing in human monocyte-derived macrophages (hMDMs) shows mRNA copies of BCAT enzymes in normalized counts. n=14 healthy donor hMDMs were used for RNA-seq. (b) Chemical structure of ERG240 & leucine. (c) Enzymatic activity of recombinant human BCAT1 & BCAT2 in presence of ERG240. (d) Relative expression of IRG1, HIF1A & IL1B measured by qRT-PCR in control, ERG240-treated (20 mM, 3 h), LPS treated (100 ng ml−1, 3 h), & LPS+ERG240 treated (LPS, 100 ng ml−1; ERG240, 20 mM for 3 h) hMDMs. ns, non-significant. All expression values are normalized to those obtained for HPRT gene expression. n=3 healthy donor hMDMs were used in each group. (e) Western blot analysis of IRG1, IL-1 beta , HIF-1 alpha & beta-actin (ACTB) in control (untreated), ERG240-treated (20 mM, 3 h), LPS treated (100 ng ml−1, 3 h) & LPS+ERG240 treated (LPS, 100 ng ml−1; ERG240, 20 mM for 3 h) hMDMs. The experiment is representative of three independent experiments using n=3 healthy donor hMDMs each. (f) GC/MS results showing itaconic acid production in hMDMs in presence or absence of ERG240 (20 mM). LPS treatment was for 8 h (100 ng ml−1). GC/MS plot for each donor is shown separately where itaconate abundance denotes arbitrary units. Error bars are s.e.m. Significance was tested using two-tailed Student’s t-test or one-way ANOVA. * P<10−3 following two-way ANOVA. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms16040), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody [NB100-449] - HIF-1 alpha /2 alpha expression in myeloid-specific KO mice targeting the HIF pathway. (A) Images of the colon of wild-type (WT), myeloid-specific Hif-1a KO (hMRP8 Hif-1a KO) or von Hippel Lindau (Vhl) KO (hMRP8 Vhl KO) mice, immunostained for MRP8 (green) & the DNA-binding regions of Hif-1a mRNA (red). Mice were fed with 5% DSS for 4 days prior to immunostaining analyses. Note that there were no MRP8-positive cells that were positive for Hif-1a mRNA in hMRP8 Hif-1a KO (middle column) mice, but we observed many cells that were double positive for MRP8 & Hif-1a mRNA in hMRP8 Vhl KO mice (right column). (B) Images of the colon of hMRP8 Vhl KO mice fed with 5% DSS as in A, immunostained for MRP8 (green) & HIF-1 alpha (red, upper row) or HIF-2 alpha (red, bottom row). DAPI-stained nuclei are shown in blue. White boxes in A & B indicate the regions magnified in the lower or right images, respectively. Yellow arrowheads in A & B indicate cells positive for both markers. Scale bars: 100 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29967068), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody [NB100-449] - Increased infiltration of myeloid cells expressing HIF-1 alpha in DSS-induced colitis. (A) Western blot analysis of MRP8, F4/80 or actin using the whole colon tissue lysate from mice fed with water or 5% DSS for 4 days. (B) Images of the colon of mice fed with water or 5% DSS, immunostained for myeloid cells using anti-MRP8 (green) or anti-HIF-1 alpha (red) antibodies. Nuclei were counterstained with DAPI. White rectangles indicate the areas magnified in the images shown below. Yellow arrowheads indicate cells with colocalization of MRP8 & HIF-1 alpha . Quantification of MRP8-positive cells & percentage of those expressing HIF-1 alpha are shown as bar graphs on the right-hand side. Data are mean±s.e.m. for at least three independent fields examined per mouse (n≥3 per group). **P<0.01 & ***P<0.001, assessed by Student's t-test. HPF, high-powered field. (C) Images of the colon of mice fed with 5% DSS, immunostained for MRP8 (red) & CD11b (green). DAPI-stained nuclei are shown in blue & a merged image is also shown. Scale bars: 100 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29967068), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Increased HIF-1 alpha expression is regulated by TSLPR inhibition & not IL-1 beta secretion (A) Human mDC were stimulated with SC glucan with either anti-TSLPR or IgG isotype control antibodies for 8 h in the presence or absence of IL-1 beta neutralization antibodies (n = 1 representative donor presented, three separate experiments performed). Pro-IL-1 beta , IL-1 beta , HIF-1 alpha , phospho-p38 MAPK, p38 MAPK, phospho-AMPK, AMPK & beta -actin were measured by immunoblot. (B,C) Densitometry of cumulative data was performed using Image Studio Lite software with HIF-1 alpha normalized to beta -actin & phospho-AMPK normalized to AMPK. Data is reported as percentage of maximal signal observed within each donor (n = 3 independent donors, presented as pooled data). Cumulative data displayed as mean +SEM. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31139177), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - miR-18b (miR-18b-5p) regulates Hif1 alpha & reduces apoptosis in mtNSC-34 cells. a Overexpressed miR-18b (miR-18b-5p) decreased Hif1 alpha & Mef2c proteins. Both Mctp1 & Rarb expression were increased by miR-18b (miR-18b-5p). Downregulated Bax & upregulated Bcl2 by miR-18b (miR-18b-5p) diminished apoptosis in mtNSC-34 cells. b RT-qPCR analysis showed low expression of Hif1 alpha & Mef2c mRNAs. c Mctp1 & Rarb transcripts were highly expressed by miR-18b (miR-18b-5p). d Bax mRNAs were decreased & Bcl2 mRNAs were increased by overexpressed miR-18b. e miR-18b (miR-18b-5p) was overexpressed in mtNSC-34 cells. f miR-206 was reduced by miR-18b (miR-18b-5p). g LDH release analysis explained that transfected miR-18b (miR-18b-5p) restores apoptosis. h Luciferases assay with 3′ UTR of Hif1 alpha showed that Hif1 alpha is target of miR-18b in contNSC-34 cells. i & j Overexpression of miR-18b (miR-18b-5p) enhanced neuronal differentiation (MAP2) & attenuated intracellular Ca2+ levels (Cont (0.098) versus miR-18b (miR-18b-5p) (0.051) in fluorescence intensities from baseline 490/525 ratio) in mtNSC-34 cells. Empty vector served as a negative control (Cont). Arrow represents SOD1 aggregation (green). Scale bar, 40 μm. Significantly different at *, p < 0.05; **, p < 0.005. The experiments were replicated 5 times Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32605607), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Hypoxic conditions upregulate CNN3 in BeWo cells.BeWo cells were cultured either under normoxic or under hypoxic conditions for 24(A) Total protein lysates were examined with the Western Blot technique to detect CNN3 protein levels. For normalization, HPRT was stained on the same membrane. As hypoxia marker, HIF-1 alpha was detected as well. (B) Protein bands of CNN3 & HPRT were densitometrically measured on the Western Blot membrane & CNN3/HPRT levels are plotted in the graph (white column: normoxia; black column: hypoxia). n = 3. (C) BeWo cells were serum starved for 16 h & then treated with either PBS as control or 200 µM CoCl2 for 6 h in serum free medium. Then total protein was isolated & a Western Blot was performed. The HIF-1 alpha, CNN3 & HPRT protein was detected on the membrane with specific antibodies. (D) A densitometric analysis was performed to determine CNN3/HPRT levels (white column: normoxia; black column: hypoxia). n = 3. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25050546), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Dynamics of the hypoxia-triggered decrease in MET phosphorylation & its reversal upon reoxygenation(A) MCF10A cells were incubated under normoxia or hypoxia for 1 h. They were then treated, under the same oxygen pressure, with 10 ng/mL HGF/SF for 5, 10, 20, 30, 60, 90 & 120 minutes. A control (Ctrl) without any HGF/SF stimulation was also performed. The same amount of protein was analyzed by western blotting with antibodies directed against: phosphorylated residues in the MET kinase domain, the MET kinase domain, the hypoxia marker HIF1a, phosphorylated Akt, Akt, Erk2, phosphorylated Erk, & actine. The positions of prestained molecular weight markers are indicated. Arrows indicate the positions of precursor & mature full-length MET. (B) MCF10A cells were incubated under hypoxia for 5, 10, 15 or 30 minutes. Another set of cells were incubated under hypoxia for 1 h & then returned to normoxia for 5, 10, 15 or 30 min (re-oxygenation). A control under normoxic (N) conditions was also included. The same amount of protein was analyzed by western blotting as previously described with the addition of GAB1 & its phosphorylated form. (C) MCF10A cells were placed under normoxic or hypoxic conditions for 1 h or hypoxia for 1 h & then normoxia for 10 minutes (reoxygenation). Cells were then treated at the indicated time with 10 ng/mL of HGF/SF. Cell lysates were incubated for AlphaScreen specific phospho-Erk & phospho-Akt quantitation. Error bars represent standard deviations (n = 3; ± SD). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29930749), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - The role of HIF-1 in purinosome formation.a, quantifying the number of purinosome-containing cells in normoxia or hypoxia (24 h) in purine-rich medium & normoxia in purine-depleted medium (purine -ve), cells in hypoxia transfected with siRNA to HIF-1 alpha (+ siRNA), & cells in purine-rich medium supplemented with DFX. Data shown are n = 3, mean ± S.E., total number of cells counted are shown in parentheses. b, time course of purinosome formation in hypoxia shows the number of purinosome-containing cells steadily increases after 3 h in hypoxia. Re-oxygenation of the samples after hypoxic incubation for 10 h reverts the number purinosome-containing cells back to normoxic levels. Data shown is n = 3, mean ± S.E., total number of cells counted are shown in parentheses. c, time course of HIF-1 alpha stabilization in hypoxia shows maximum HIF-1 alpha protein expression levels at 3 h in hypoxia, after which the HIF-1 alpha expression decreases. The positions of molecular markers are shown for each blot; uncropped blots with overlaid markers are deposited in the raw data files. d, the effect of hypoxia on the transcription of purine biosynthesis enzymes measured by qPCR. Vascular endothelial growth factor (VEGF) & HIF-1 alpha are controls. Data shown are n = 5, mean ± S.E. e, the effect of hypoxia on the protein expression levels of the purine biosynthetic enzymes. HIF-1 alpha is stabilized in hypoxia as expected, & no significant increase in the purine enzymes was detected between normoxic (21% oxygen) & hypoxic (1% oxygen) growth conditions. The positions of molecular markers surrounding each band of interest are shown for each blot; uncropped blots with overlaid markers are deposited in the raw data files. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32439803), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - C/EBPδ promotes HIF-1 alpha expression in macrophages through inhibition of FBXW7 alpha a) RT-PCR analysis of FBXW7 isoform expression from different sources as follows. 1, primary peritoneal macrophages (PPMs); 2, RAW 264.7 cells; 3, MMTV-Neu mammary tumour tissue; 4, primary mouse embryo fibroblasts. Numbers indicate the position of size markers in base pairs. (b) RT-qPCR analysis of Fbxw7 transcript levels in PPMs from WT & Cebpd−/− KO mice, cultured +/− LPS (100 ng/ml, 24 h), compared to WT untreated (n=4, *P<0.05; **P<0.001). (c) Western analysis of nuclear extract (NE) from primary human monocytes nucleofected with siRNA oligos (C, control; D, CEBPD; F, FBXW7) & treated with LPS (100 ng/ml) & 1%O2 (16 h) as indicated. SE, short exposure; LE, long exposure. (d) RT-qPCR analysis of FBXW7 & CEBPD transcripts in primary human monocytes as in panel (c) (n=3, *P<0.05; **P<0.001). (e) Western analysis of NE from PPMs nucelofected with siRNA oligos & treated with LPS (100 ng/ml) & 1%O2 for 16 h as indicated. SE, short exposure; LE, long exposure.Where applicable, data are mean±S.E.M., evaluated by two-tailed unequal variance t-test. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23575666), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Hypoxia downregulates CFTR expression. (a) Phase-contrast photographs of MDCK cells treated with normoxia & hypoxia conditions; (b) Western blot showing downregulation of CFTR in the MDCK cells by hypoxia (Full-length blot is shown in Supplementary Figure S7b.); (c) Real time-PCR assay showing decreased mRNA expression of CFTR induced by hypoxia in HK-2 cells,*p < 0.05; (d) Western blot showing decreased expression of CFTR induced by hypoxia in HK-2 cells (Full-length blot is shown in Supplementary Figure S7c.); (e) Western blot showing the expression changes of HIF-1 alpha & EMT markers induced by hypoxia in MDCK cells, quantification analysis is shown in the lower panel, *p < 0.05; (Full-length blot is shown in Supplementary Figure S7d.) (f) Immunofluorescent staining showing dramatically increased HIF-1 alpha & reduced CFTR protein levels in tubular epithelial cells at inner cortices in UUO kidney (arrow). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28701694), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - NRP1 & MET on HT1080 cells are accessible for blood-borne rhodocetin-alpha beta (A) expression of NRP1 & MET by HT1080 cells was proven by flow cytometry. Gray, isotype-matched controls. (B) flow cytometry of NRP1-knockout HT1080 cells demonstrating their NRP1-deficiency & unaffected MET expression. (C) treatment with CoCl2, mimicking a hypoxic tumor micro-environment, induced upregulation of HIF-1 alpha . (D) NRP1, as a downstream target of HIF-1 alpha , is upregulated in HT1080 cells but not in NRP1-knockout HT1080 cells. beta -actin immunoblots show even loading. (E) increased HIF-1 alpha (red) levels in hypoxic tumor regions, which also contained partly (arrows) or completely (open arrows) EC-deficient VM vessels. ECs are stained green & nuclei blue. (F) immunostaining of NRP1 (red) & MET (blue) showed that both proteins were present on HT1080 cells & ECs in tumor tissue. Note the continuity between EC-lined vasculature & EC marker-deficient vessels (arrows in F’). F’, F’’, in oblique view, gating of the green CD31 signal also showed an apical absence of NRP1 on ECs in contrast to MET (open arrows). (G) the fluorescence intensity along a traceroute, averaged over a width of 5 pixels, (rectangle in F) through the endothelium revealed that in ECs NRP1, unlike MET, is absent from the apical side & restricted to the basolateral side. In contrast, on ATV/VM-lining cells (F’, arrows) both NRP1 & MET are accessible from the bloodstream (H). Vertical gray lines in G & H indicate the position of the apical cell border. Original magnification was 400× (E) & 630× (F-F’’). Representative images are shown. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.25032), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody [NB100-449] - NRP1 & MET on HT1080 cells are accessible for blood-borne rhodocetin-alpha beta (A) expression of NRP1 & MET by HT1080 cells was proven by flow cytometry. Gray, isotype-matched controls. (B) flow cytometry of NRP1-knockout HT1080 cells demonstrating their NRP1-deficiency & unaffected MET expression. (C) treatment with CoCl2, mimicking a hypoxic tumor micro-environment, induced upregulation of HIF-1 alpha . (D) NRP1, as a downstream target of HIF-1 alpha , is upregulated in HT1080 cells but not in NRP1-knockout HT1080 cells. beta -actin immunoblots show even loading. (E) increased HIF-1 alpha (red) levels in hypoxic tumor regions, which also contained partly (arrows) or completely (open arrows) EC-deficient VM vessels. ECs are stained green & nuclei blue. (F) immunostaining of NRP1 (red) & MET (blue) showed that both proteins were present on HT1080 cells & ECs in tumor tissue. Note the continuity between EC-lined vasculature & EC marker-deficient vessels (arrows in F’). F’, F’’, in oblique view, gating of the green CD31 signal also showed an apical absence of NRP1 on ECs in contrast to MET (open arrows). (G) the fluorescence intensity along a traceroute, averaged over a width of 5 pixels, (rectangle in F) through the endothelium revealed that in ECs NRP1, unlike MET, is absent from the apical side & restricted to the basolateral side. In contrast, on ATV/VM-lining cells (F’, arrows) both NRP1 & MET are accessible from the bloodstream (H). Vertical gray lines in G & H indicate the position of the apical cell border. Original magnification was 400× (E) & 630× (F-F’’). Representative images are shown. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.25032), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Dectin-1-induced TSLP negatively regulates pro-IL-1 beta & HIF-1 alpha . (A) Human mDC were stimulated with SC glucan, CA glucan or heat killed C. albicans hyphae with anti-TSLPR antibodies or IgG isotype control for 24 h (n = 6 independent donors, presented as pooled data). Lactate production was measured in cell-culture supernatants using colourmetric L-lactate detection kit. (B) Human mDC were stimulated SC glucan with either anti-TSLP, anti-TSLPR or IgG isotype control antibodies for 8 h (n = 1 representative donor presented, three separate experiments performed). Pro-IL-1 beta , IL-1 beta , HIF-1 alpha , phospho-p38 MAPK, p38 MAPK, phospho-AMPK, AMPK & beta -actin were measured by immunoblot. (C–G) Densitometry of cumulative data was performed using Image Studio Lite software with pro-IL-1 beta , IL-1 beta & HIF-1 alpha normalized to beta -actin & phospho-p38 MAPK & phospho-AMPK normalized to total p38 MAPK & AMPK respectively. Data is reported as percentage of maximal signal observed within each donor (n = 3 independent donors, presented as pooled data). Cumulative data displayed as mean +SEM. Statistical analysis calculated using one-way ANOVA with Bonferroni post-tests (***p = 0.001). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31139177), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - Involvement of MET in Akt & Erk pathway activation under hypoxiaMCF10A cells were transfected with a pool of three MET-targeting siRNAs (20 nM) or a control siRNA (siCtrl). A control without siRNA was also included (Ctrl) (A). MCF10A cells were transfected with two HIF1a-targeting siRNAs (20 nM), independently or together, or a control siRNA (siCtrl) (B). The cells were then placed for 1 h under normoxic or hypoxic conditions & treated or not for 10 min with 10 ng/mL HGF/SF. In each experiment, the same amount of protein was analyzed by western blotting with antibodies directed against: phosphorylated residues in the MET kinase domain, the MET kinase domain, phosphorylated Akt, Akt, phosphorylated Erk, Erk2, phosphorylated GAB1, GAB1, or hypoxia marker HIF1a. The positions of prestained molecular weight markers are indicated. Arrows indicate the positions of precursor & mature full-length MET & Erk1/2 proteins. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29930749), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody [NB100-449] - beta -Glucan size affects the requirement for reactive oxygen species (ROS) in IL-1 beta induction. (A) Human monocyte-derived dendritic cell (mDC) stimulated with curdlan or glucan-mp for 8 h. ROS were detected by incubating cells with CellRox Green fluorescence dye & analysis by flow cytometry (representative experiment of two) presented, (B) Human mDCs from healthy donors or chronic granulomatous disease (CGD) patients were stimulated with curdlan or glucan-mp for 8 h. HIF-1 alpha & pro-IL-1 beta protein expression measured by immunoblot (n = 2 donors). (C–L) Human mDC from healthy donors or CGD patients were stimulated with curdlan or glucan-mp for 24 h (n = 6 donors). IL-1 beta , IL-6, IL-23, TSLP, & CCL22 secretion were measured by enzyme-linked immunosorbent assay, with cumulative data displayed as mean ± SEM. Image collected & cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fimmu.2017.00791/full), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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

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NB100-134

HIF-1 alpha Antibody - BSA Free

(14 Reviews)

(236 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).

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 & are expressed relative to control (n = 3). (B,C) Increased Wnt11 mRNA by hypoxia in EMSC preadipocytes & adipocytes (Day 0–12 after differentiation) (B), & C2C12 myoblast & myocyte (Day 0 & 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 & 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) & 4T1 cells (F). (G) EMSCs treated with 0.1 mM DMOG for the indicated times. Wnt11 & Vegf mRNA expression was measured by qPCR & 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 25 pictures/group from three separate experiments. Scale bar = 40 µm. (F) Cell proliferation was assessed by evaluating PCNA expression, n = 4. * p 20). B May–Grunwald–Giemsa staining of whole KM blood cells in 6-month-old AB zebrafish after 4 days of E2 treatment (t-test, ***p 20). F Effect of E2 on neutrophil proliferation in AB zebrafish embryos. (one-way ANOVA (LSD). ns, no significance, n > 10). G E2 promotes the apoptosis of myeloid lineage cells in AB zebrafish embryos (one-way ANOVA (LSD) **p 10). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35842445), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-134] - IGFBP5 inhibits HIF1 alpha expression through the MAPK-ERK signaling pathwayA. & B. Western blots using specific antibodies for the phosphorylation state of IGF1R, ERK1/2, & p38-MAPK from A375 IGFBP5 OE & control cells. Phosphorylation of IGF1R, ERK1/2, & p38-MAPK was decreased in A375 cells transfected with IGFBP5. GAPDH was used as a loading control. C. Overexpression of IGFBP5 inhibited HIF1 alpha expression visualized by western blots of A375 cells. The arrow points to the band of HIF1 alpha . D. Assessment of the reduced gene expression levels of VEGFA & MMP9, downstream genes regulated by HIF1 alpha , 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 & vector control cells. *, P < 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26010068), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-134] - WNT11 regulates MMPs activities.(A–C) (Top panels): Serum-free medium was conditioned for 24 hrs by the indicated cells, concentrated 20-fold & assayed by gelatin zymography. Gelatinolytic activity is indicated by clear zones against a dark background of stained substrate. (Bottom): Whole cell extracts were immunoblotted with indicated antibodies. (A) Overexpression of Wnt11 in EMSC or BT473 cells enhances activity of MMP-9 & MMP-2. (B) Impaired activity of MMP-9 & MMP-2 in MDA-MB-231 cells (left) or EMSCs (right) stably expressing Wnt11 shRNAs & treated with DMOG. (C) WNT11 is required for MMP-9 & MMP-2 activity in MDA-MB-231 cells (left) or EMSCs (right) under normoxic & hypoxic culture conditions. (D) WNT11 regulates MMP2 protein in media. (Top): conditioned media from indicated cells & treatments. (Bottom): whole cell lysates were immunoblotted with indicated antibodies. (E) Recombinant WNT11 induces both MMP-2 protein & MMP-2 activity in media. (Top panels): Gelatin zymography & immunoblot of serum-free medium conditioned for the indicated times after recombinant WNT11 (r-WNT11) treatment. (Bottom): Whole cell lysates were immunoblotted with indicated antibodies. (F) MMP-2 inhibitor attenuated induced migration by WNT11. MDA-MB-231 cells infected with lentiviruses for stable expression of Wnt11 or GFP (n = 4) were incubated with either vehicle or 1 μM of ARP100. Media in the lower compartment had same concentration of DMSO or inhibitor. Values are mean ± s.e.m. *p < 0.05, **p < 0.01. For panels (A–D), HIF-1 alpha & HIF-2 alpha were shown as a marker of hypoxia, WNT11 normalized to alpha -Tubulin was shown. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-134] - ER-INP binds to PHD2, inhibits hydroxylation of HIF-1 alpha & increases HIF-1 alpha accumulation. (A) Co-immunoprecipitation assay. HEK293 cells were transfected with control vector or intrabody ER-INP for 48 hours & subsequently lysed. Co-immunoprecipitation & western blot assays were performed on the cell lysis. ER-INP recognized & bound to PHD2 in HEK293 cells (n = 3). (B) Western blot analysis to measure the effect of ER-INP on HIF-1 alpha & its hydroxylation level in transfected HEK293 & RAW264.7 cells pre-treated with MG132 (upper panel) & the protein ratio to beta -actin loading control by ImageJ densitometry analysis (lower panel; n = 3). (C) Immunofluorescence assay. Expression of ER-INP increases HIF-1 alpha accumulation in RAW264.7 cells. Cells were stained with anti-HIF-1 alpha antibody & DAPI & then visualized & photographed under immunofluorescence microscopy (left), & mean fluorescence intensity of HIF-1 alpha versus the mean fluorescence intensity of nuclear DAPI staining is displayed (right). Data represent the mean ± SD of 3 independent slides. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31413262), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-134] - HIF-1 alpha binds to TIM-3 promoter & regulates its expression in primary glia.(a) Cell surface expression of TIM-3 analysed in BV2 cells under 20% O2 or 1% O2 for 24 h by flow cytometry using PE-conjugated anti-TIM-3 antibody. Results from 3 independent experiments presented as a representative histogram & the mean fold change (± s.d.) relative to normoxic sample. (b) Mouse primary mixed glial cells incubated under hypoxia or normoxia for 24 h, & the cells examined by immunocytochemistry using an anti-TIM-3 antibody. (c,d) Mouse primary mixed glial cells & primary neuronal cells incubated under hypoxia or normoxia for 24 h, & then RT–PCR used to detect the levels of TIM-3 & actin. Relative transcript levels shown as the mean fold change (± s.d.) from 3 independent experiments (NS, not significant, Student–Newman–Keuls test). (e) Primary mixed glial cells incubated under hypoxia or normoxia for 24 h, & chromatin immunoprecipitation (ChIP) performed w/ anti-HIF-1 alpha or control IgG. Results presented as relative amounts representative of 3 independent experiments. (f) Primary mixed glial cells cultured from HIF-1 alpha +f/+f mice, infected w/ Ad-GFP or Ad-Cre/GFP, transfected w/ TIM-3-luciferase reporter constructs & 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) & WB analysis (h) performed under hypoxia or normoxia for 24 h using the indicated primers & antibodies, respectively. The data shown representative of at least 3 independent experiments. The graphs show the % changes in TIM-3 transcript & protein levels in Ad-Cre/GFP- versus Ad-GFP-infected cells under hypoxia. IP, immunoprecipitation. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25790768), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-134] - Regulation of catabolic genes by HIF-1 alpha . (A) Protein levels of Hif1a, & mRNA levels of Mmp13, & Hif2a in WT mouse primary chondrocytes transduced with GFP or HIF1A adenoviral vectors under the hypoxic condition (3% O2). GFP or HIF1A was transduced at a multiplicity of infection (MOI) of 100. The cells were treated with or without 10 ng/mL IL-1 beta for 2 days. Bars show the mean ± SD of three samples per group. *P < 0.05. (B) Protein levels of Hif1a, & mRNA levels of Mmp13, & Hif2a in WT mouse primary chondrocytes transfected with siRNA against GFP or Hif1a under the hypoxic condition. The cells were treated with or without 10 ng/mL IL-1 beta for 2 days. Bars show the mean ± SD of three samples per group. *P < 0.05. (C) Protein levels of Hif1a, & mRNA levels of Mmp13, & Hif2a in Hif1afl/fl primary chondrocytes transduced with GFP or Cre adenoviral vectors under the hypoxic condition. GFP or Cre was transduced at a MOI of 100. The cells were treated with or without 10 ng/mL IL-1 beta for 2 days. Bars show the mean ± SD of three samples per group. *P < 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32214220), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-134] - NSCs Reduce Succinate Levels & Reprogram the Metabolism of Type 1 Pro-inflammatory Mφ toward Oxidative Phosphorylation In Vitro(A) Experimental setup for in vitro MφLPS co-cultures with iNSCs/NSCs.(B & C) Gene expression microarrays of MφLPS-iNSCs/NSCs. (B) Venn diagram of differentially expressed genes (adjusted p value < 0.1). (C) Heatmap of genes differentially expressed (adjusted p value < 0.1) in MφLPS-iNSCs or MφLPS-NSCs.(D & E) qRT-PCR independent validation of differentially expressed inflammatory genes as in (C). (D) Expression of genes related to type 1 inflammatory (E) & anti-inflammatory Mφ phenotypes relative to Actb. Data are mean fold change (±SEM) versus MφLPS from n ≥ 3 independent replicates per condition.(F) qRT-PCR of BV2LPS-iNSCs/NSCs (±SEM) from n ≥ 3 independent experiments per condition. BV2 & BV2LPS are shown as controls.(G & H) Extracellular flux (XF) assay of the oxygen consumption rate (OCR) (G) & extracellular acidification rate (ECAR) (H) in MφLPS-iNSCs/NSCs. Data were normalized on total protein content & are expressed as mean values (±SEM) from n ≥ 3 independent experiments per condition.(I & J) Levels of significantly changed extracellular (EXTRA_Metab, I) & intracellular (INTRA_Metab, J) metabolites in MφLPS versus Mφ at 25 hr. Data are mean a.u. (±SEM) from n ≥ 2 independent experiments per condition.(K & L) Hif-1 alpha (K), PKM2 (K), & IL-1 beta (L) expression levels relative to beta -actin. Data are mean fold change versus MφLPS (±SEM) from n ≥ 3 independent experiments per condition.∗p ≤ 0.05 & ∗∗p ≤ 0.01 versus MφLPS. See also Tables S2 & S3. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29478844), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody - BSA Free [NB100-134] - Enhanced expression of hypoxia-induced survival factors & angiogenic growth factors in hASC L-spheroids.(A) The light source used was LED (660 nm) designed to fit over a microplate (12.5 × 8.5 cm) for cell culture. (B) Formation of hASC L-spheroids. hASCs morphology on non–tissue culture–treated 24-well plates at day 3. Scale bar = 500 μm. (C) Western blot analysis & quantification of HIF1-alpha in hASCs cultured as spheroids, L-spheroids & monolayers (*p < 0.01, compared to the L-spheroid group). (D) Angiogenesis-related protein analysis of L-spheroids (*, p < 0.05, compared to the spheroid group, t-test, n = 3 in each group). (E) ELISA measurement of spheroids cultured for 3 days. Concentrations of VEGF are presented as pg-corrected for 104 cells. (*, p < 0.05, compared with spheroid 6J/cm2 group, t-test, n = 3 in each group). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26065900), 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
CoraFluor 1	NB100-134CL1
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 525	NB100-134JF525
Janelia Fluor 549	NB100-134JF549
Janelia Fluor 646	NB100-134JF646
Janelia Fluor 669	NB100-134JF669
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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NB100-123

HIF-1 alpha Antibody (H1alpha67)

(9 Reviews)

(165 Publications)

0.1 ml

NB100-123

<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - MLN4924 induces accumulation of HIF1a in a time dependent manner. Cells were treated with 0.1uM MLN4924 for indicated time periods, followed by IB with indicated antibodies. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2012125) licensed under a CC-BY license.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Analysis of HIF-1 alpha in human lung tissue. Image courtesy of product review by Aneta Gandjeva.

Immunoprecipitation: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - MG132 attenuates the inhibitory effect of interleukin-1beta on HIF-1/AM axis. U87MG cells were incubated for 2 h in hypoxia (1% O2) with or without interleukin-1beta (10 ng/ml). The proteasome inhibitor MG132 was added 30 min in advance to prevent ubiquitinated HIF-1alpha from proteasomal degradation. HIF-1alpha was precipitated by anti-HIF-1alpha antibody. Ubiquitin was detected by immunoblotting. Non-specific mouse IgG was used as a negative control. The results are representative for three independent experiments Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2013562) licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Analysis using the HRP conjugate of NB100-123. Detection of HIF-1 alpha in cobalt chloride treated/untreated COS-7 nuclear extracts using NB100-123.

$<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Analysis using the HRP conjugate of NB100-123. On day 1, MEF cells (+/+,-/-), were grown on 15cm dish (2x10 to the 6th cells). On day 2, cells were exposed to hypoxia for 4hrs. Cells were washed with ice cold PBS twice and whole cell protein was extracted extracted with RIPA buffer fortified with protease. Upon quantification, 100ug of protein was fractionated on 7% polyacralymide gel. Gel was transferred overnight onto nitrocellulose membrane. The membrane was probed with HIF-1 alpha monoclonal antibody at a 1:500 dilution (NB100-123). The secondary antibody was conjugated with HRP and was used at a 1:2500 dilution. Photo courtesy of Dr. Gregg Semenza, Johns Hopkins University.$

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Dapagliflozin protects against metabolic switch by inhibition of HIF-1 alpha. Representative images of PTCs incubated with 10 umol/l of molidustat for 6 h with immunofluorescent staining for HIF-1 alpha. Scale bar = 10 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33187129/) licensed under a CC-BY license.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Immunohistochemical detection of HIF-1alpha protein in human tissues. The nature of the tissue is indicated on top of each figure. Original magnifications are as follows: A, x400; B, x400; C, x100; D, x100; E, x100; F, x100; G, x40; H, x40. Image collected and cropped by CiteAb from the following publication (//bmcgenet.biomedcentral.com/articles/10.1186/1471-2156-5-27), licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Staining in pig endothelial cells under hypoxia condition using NB100-123. Image from verified customer review.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Staining on pig tissue (brown) using NB100-123. Image from verified customer review.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Staining of biotin conjugated HIF-1 alpha (NB100-123B) in human kidney.

Immunoprecipitation: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Immunoprecipitation of HIF-1alpha using NB100-123 in COS7 CoCl2 treated lysate. Heavy and light chains are also detected. Image using the HRP form of this antibody.

Immunohistological and western blot analyses of HIF-1 alpha .(A) PAB liver stained by HIF-1 alpha (original magnification, ×20). (B) Sham-operated control liver stained by HIF-1 alpha (original magnification, ×20). (C) Enlarged view of the boxed area in the Image (A) (original magnification, ×100). (D) Western blot analysis of HIF-1 alpha (mean ± SEM), * P<0.05. HIF-1 alpha : hypoxia-inducible factor-1 alpha ; PAB: pulmonary artery banding.

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: Mouse Monoclonal HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Images of HIF-1 alpha localization (red) in CTR and CoCl2-treated oAECs after 24 h of culture. Image from a verified customer review.

Western Blot: Mouse Monoclonal HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Image of HIF-1 alpha activation in AECs control cells (CTR) and after 24h of 10 and 50µM of CoCl2-treatment; HIF-1 alpha time course at shorter CoCl2 incubation times. Image from a verified customer review.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - 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 & cropped by CiteAb from the following publication (//bmcgenet.biomedcentral.com/articles/10.1186/1471-2156-5-27), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Immunohistological & western blot analyses of HIF-1 alpha .(A) PAB liver stained by HIF-1 alpha (original magnification, ×20). (B) Sham-operated control liver stained by HIF-1 alpha (original magnification, ×20). (C) Enlarged view of the boxed area in the Image (A) (original magnification, ×100). (D) Western blot analysis of HIF-1 alpha (mean ± SEM), * P<0.05. HIF-1 alpha : hypoxia-inducible factor-1 alpha ; PAB: pulmonary artery banding. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26863419), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Analysis of CBF treatment in HCT116 implanted mouse models & tumour tissue samples. (a) Tumour size versus CBF treatment. The lowest tumour growth rate was found in i.p. group during CBF treatment. (b) Analysis of HIF-1 alpha mRNA level in tumour tissues. There was a significant upregulated HIF-1 alpha level in i.p. group. Results are means with standard errors from 4 replicates. (c) Inhibition of HIF-1 alpha nuclear translocation in the i.t. sample. CBF strongly inhibited HIF-1 alpha nuclear accumulation in the tumour of xenografts implanted with HCT116 cells. Scale bars equal 10 μm. White arrows: HIF-1 alpha in the cytosol. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23818933), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - CycT alleviates tumor hypoxia. The effects of CycT on the levels of exogenous hypoxia-marker pimonidazole labeling (A), the levels of hypoxia-inducible CA9 enzyme (B), & the levels of hypoxia-inducible factor HIF1 alpha (C) in orthotopic tumor xenografts. Scale bar: Montage, 1 mm; 10X, 20 μm. Data are plotted as mean ± SEM. For statistical analysis, the levels in treated tumors were compared to the levels in control tumors with a Welch 2-sample t-test. **p-value < 0.005. IHC images are representative of 3 independent experiments. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30723259), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Schematic demonstration of the three HIF1 alpha antibodies used in this study & their epitopes (Figs. 2a & b). Ab1 is a polyclonal antibody raised against N-domain of wild type HIF1 alpha & does not recognize HIF1 alpha 1.2 due to it's different N-terminal part (Fig. 2a). Ab2 & Ab3 are monoclonal & polyclonal antibodies, respectively, with epitopes in the common parts of HIF1 alpha & HIF1 alpha 1.2 (Fig.2a & b). Immunohistochemical analysis performed on thin adjacent sections of NE-differentiated prostate cancer using the HIF1 alpha antibodies Ab1 (Fig. 2c), Ab 2 (Fig. 2d), Ab 3 (Fig. 2e) & HIF1 beta antibody (Fig. 2f). Immunopositivity was detected for HIF1 alpha with Ab2 & Ab3 while HIF1 alpha Ab1 produced no detectable staining. HIF1 beta was also positive in adjacent section (Fig. 2f). Double staining of chromogranin A & androgen receptor antigens on adjacent sections (Fig. 2g) showed immunopositivity for chromogranin A (Fast red). Androgen receptor antibody (DAB, brown) produced no staining. Immunostaining of benign prostate tissue with HIF1 alpha Ab3 showed immunopositivity in NE-like cells of benign prostate tissue (Fig. 2h). In addition, HIF1 beta antibody recognized benign NE-like cells in benign prostate hyperplasia (Fig. 2i). Double staining with HIF1 alpha Ab3 & HIF1 beta (Fig. 2j) shows co-localization of the two proteins in NE-like cells of benign prostate hyperplasia (HIF1 alpha Ab3 red stain, HIF1 beta brown stain). Panels a, b, c, d, e, f & g: 40× objective. Panels h, i & j: 60× objective. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/20663134), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - 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) & 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 & cropped by CiteAb from the following publication (//actaneurocomms.biomedcentral.com/articles/10.1186/2051-5960-2-51), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Levels of GSH in cortical neurons exposed to hypoxia. A) During hypoxia, GSH increased in MAP2-ir neurons with round somata (bottom panel, open arrow) & decreased in neurons with pyramidal-like morphology (upper panel, solid arrow). B) GSH increased in a subset of GAD65/67–ir neurons exposed to hypoxia (bottom panel, open arrow). C) Elevated HIF-1 alpha expression in GAD65/67–ir neurons containing high levels of GSH when exposed to hypoxia (middle panel). BSO treatment decreased HIF-1 alpha expression in GAD65/67-positive neurons during hypoxia. D) Percentages of HIF-1 alpha -expressing GAD65/67-ir neurons expressing a high level of GSH & GAD65/67-ir neurons expressing a low levels of GSH & HIF-1 alpha (mean ± SD; n = 3). The “low” level referred to the level of GSH in normoxic GAD65/67 neurons that was normalized to 1. The “high” level referred to the elevated GSH level in GAD65/67 neurons with a mean of 1.40 ± 0.10. E) Total MCB-GSH intensity in GAD65/67-positive neurons with & without BSO after hypoxia (mean ± SD; 8-10 neurons quantified from each experiment, n = 3 independent cultures). Scale bars, 20 μm. Image collected & cropped by CiteAb from the following publication (//actaneurocomms.biomedcentral.com/articles/10.1186/2051-5960-2-51), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Results of in situ hybridization & immunohistochemistry on thin adjacent section to detect expression of HIF-1 alpha 1.2 mRNA & HIF1 alpha protein in malignant & benign prostate tissue. In situ hybridization (antisense probe, Fig. 3a) & immunostaining with HIF1 alpha Ab2 (Fig. 3c) on thin adjacent sections of NE-differentiated prostate adenocarcinoma showed co-localization of HIF1 alpha 1.2 transcript & HIF-1 alpha protein. Incubation with sense probe did not generate any detectable hybridization signals (Fig. 3b). Both In situ hybridization (Fig. 3d antisense) & HIF-1 alpha Ab2 immunostaining (Fig. 3f) were negative in non-NE-differentiated prostate adenocarcinoma. In situ hybridization with sense probe performed on non-NE-differentiated prostate cancer (Fig. 3e) was negative. In situ hybridizering on benign prostate tissue showed HIF1 alpha 1.2 transcript in NE-like cells of benign prostate tissue (Fig. 3g, ↗). The sense probe on thin adjacent section generated no signals (Fig. 3h,↗). Furthermore, co-localization of HIF1 alpha 1.2 transcript (Fig. 3i,↗,◢,↖) & HIF1 alpha protein, detected with HIF1 alpha Ab3 (Fig. 3j,↗,◢,↖) was also shown in NE-like cells of benign prostate tissue. Panels a, b, c, d, e, f, g, h, i, j: 40× objective. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/20663134), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - HX-induced Sirt1 expression in white matter OPCs requires HIF1 alpha .(a) HIF1 alpha stabilization of Sirt1 transcript expression in OPCs as revealed by representative RT–PCR represents higher level of Sirt1 mRNA in VHL cKO mice. GDPDH mRNA serves as a control. Mean±s.e.m., n=3 brains for each group. (b,c) Representative western blot demonstrates a transient increase of HIF1 alpha expression in HX white matter at P18 with no significant effect at P11 (P=0.7955) & P45 (P=0.7333). Histograms show mean±s.e.m. (d,e) Graphs represent the percentages of Sirt1+Ki67+ & NG2+Ki67+ cells after HX white matter in WT & HIF1 alpha KO mice. Number in parentheses within bar indicates number of samples (n=4 brains per group & per genotype; ****P<0.0001, one-way analysis of variance, Bonferroni post hoc test, mean±s.e.m.). (f,g) Western blot demonstrates no increase (P=0.8231) in Sirt1 & HIF1 alpha expression in white matter of Hif1 alpha KO mice. Actin was used as loading control (mean±s.e.m.; n=3 brains for each experiment, group & genotype). Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/ncomms13866), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - HIF-1 alpha expression in primary cortical neurons exposed to hypoxia/ischemia. A & A’) Neurons were double-stained for HIF-1 alpha & MAP2 in the presence of 5 & 25 mM glucose with & without hypoxia. HIF-1 alpha expression in the somata was observed in cells with interneuron-like morphology after hypoxia. B) CoCl2 (0.3 mM) induced HIF-1 alpha expression in cells with interneuron-like morphology. C) Quantification represents the increase in HIF-1 alpha –ir staining (mean ± SD; 5-10 neurons quantified from each experiment, n = 3 experiments). D)In vivo brain slice shows a similar pattern of positive HIF-1 alpha -ir in round soma (open arrow) & negative in neurons with pyramidal-like morphology (solid arrow) in the ipsilateral side. *p < 0.05, compared with normoxia (25 mM glucose), #p < 0.05, compared with hypoxia (25 mM glucose). Scale bars, 20 μm (A, A’, B); 10 μm (D). Image collected & cropped by CiteAb from the following publication (//actaneurocomms.biomedcentral.com/articles/10.1186/2051-5960-2-51), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Po, Av, Bv, Ca, Ma, Pm, Rb, Sh

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Av, Bv, Ca, Ma, Pm, Rb, Sh

Applications:

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

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

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-123AF350
Alexa Fluor 405	NB100-123AF405
Alexa Fluor 488	NB100-123AF488
Alexa Fluor 532	NB100-123AF532
Alexa Fluor 594	NB100-123AF594
Alexa Fluor 647	NB100-123AF647
Alexa Fluor 700	NB100-123AF700
Alexa Fluor 750	NB100-123AF750
Allophycocyanin	NB100-123APC
Biotin	NB100-123B
CoraFluor 1	NB100-123CL1
DyLight 350	NB100-123UV
DyLight 405	NB100-123V
DyLight 488	NB100-123G
DyLight 550	NB100-123R
DyLight 594	NB100-123DL594
DyLight 650	NB100-123C
DyLight 680	NB100-123FR
DyLight 755	NB100-123IR
FITC	NB100-123F
HRP	NB100-123H
Janelia Fluor 525	NB100-123JF525
Janelia Fluor 549	NB100-123JF549
Janelia Fluor 646	NB100-123JF646
Janelia Fluor 669	NB100-123JF669
PE	NB100-123PE
PerCP	NB100-123PCP
mFluor Violet 450 SE	NB100-123MFV450
mFluor Violet 500 SE	NB100-123MFV500
mFluor Violet 610 SE	NB100-123MFV610

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NB100-2284

CD31/PECAM-1 Antibody

(5 Reviews)

(151 Publications)

0.1 ml

NB100-2284

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

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

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

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

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

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

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

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

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

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

Immunohistochemistry: Rabbit Polyclonal CD31/PECAM-1 Antibody [NB100-2284] - Staining of CD31 in human lung tissue. Nuclei in blue and CD31 (vessel) in green. Mouse anti-CD31 (1:100, overnight incubation at +4°C), followed by donkey anti-mouse 488 (1:400, 3h at RT). Image from a verified customer review.

Immunohistochemistry: CD31/PECAM-1 Antibody [NB100-2284] - Antibodies selected for characterization of stromal cells & vasculature in FFPE specimens are species-specific. (A, B) Labeling of a patient colon tumor (A) & Colo205 human cell line xenograft (B) with anti-human mitochondrial antibody (single arrow- tumor cells; double arrows- stroma). (C, D) Labeling of a patient pancreatic tumor (C) & Colo205 human cell line xenograft (D) with anti-human CD34. (E, F) Labeling of a patient lung tumor (E) & Colo205 xenograft (F) with anti-human CD31. (G, H) Labeling of a patient lung tumor (G) & Colo205 xenograft (H) with anti-mouse CD34. (bars = 50 μ). Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: CD31/PECAM-1 Antibody [NB100-2284] - At the time of second passage, vessels in successfully growing xenografts of four different tumor types surveyed were of murine origin: A-C Colon; D-F Lung; G-I Pancreatic; J-L Renal Cell Carcinoma. For each tumor type, a representative section of an original patient specimen labeled with anti-huCD31 is shown (A, D, G, J). For each tumor, sections of the first passage xenografted tumor when it was resected are also shown; while no huCD31(+) vessels were identified in the xenografts (B, E, H, K), msCD34(+) vessels were abundant (C, F, I, L). (bars = 50 μ). Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: CD31/PECAM-1 Antibody [NB100-2284] - Vascularization of an engrafted patient colon tumor. A patient colon tumor was implanted in a cohort of mice & vessel development was analyzed over 8 weeks. Vessels in the original patient specimen labeled strongly for huCD31 (A) & not for msCD34 (D). Representative sections showing loss of huCD31(+) vessels (B- 4 weeks, C- 7 weeks) & presence of msCD34(+) vessels (E- 4 weeks, F- 7 weeks) are shown. The graph (G) summarizes this process; huCD31(+) vessels were rapidly lost, & by one week, mouse vessels were the predominate vessels present in the colon tumors (no data for msCD34 at 2 weeks; bars = 100 μ). Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: CD31/PECAM-1 Antibody [NB100-2284] - KCl pulse decreased membrane, but increased nuclear, detection of NHE1 in b.End3 endothelial cells.(A) Representative immunofluorescence images of bEnd.3 cells at two time-points after KCl or aCSF pulse. (B) Representative immunoblots of NHE1, alpha -tubulin, PECAM, & lamin B in cytosol, membrane, & nuclear fractions of bEnd.3 cells harvested at 5 min after KCl or aCSF pulse. (C = cytosol, M = membrane, N = nuclear) Values represent the mean ratio of NHE1 detection ± SEM (n = 11–12). (C) Representative immunoblots indicating NHE1 & alpha -tubulin as a loading control in whole cell lysate of bEnd.3 cells harvested at 5 min after KCl or aCSF pulse. Values represent the % of aCSF-treated relative expression ± SEM (n = 6). (D) Representative immunoblots of NHE1 & alpha -tubulin as a loading control in whole lysate of microvessels harvested at 90 min after cortical injection of KCl or aCSF. Values represent the % of naive relative expression ± SEM (n = 6). # denotes significantly different vs naïve (p<0.01), as assessed by one-way ANOVA (E) Intracellular pH during aCSF or KCl pulse. All data represent mean ± SEM (n = 45). (F) Extracellular pH during aCSF or KCl pulse. All data represent mean ± SEM (n = 6) in triplicate. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32469979), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Immunocytochemistry/ Immunofluorescence: CD31/PECAM-1 Antibody [NB100-2284] - Impact of T‐AuNPs on endothelial cells in vivo. Immunofluorescent staining for endothelial cells using an anti‐CD 31 antibody revealed an intact endothelial cell monolayer in the uninjured control carotid artery (N = 3). As expected post balloon angioplasty, there is no endothelial cell monolayer of the injured left carotid arteries in the injury alone, injury + T‐AuNP, & injury + S‐AuNP treatment conditions. Green indicates autofluorescence of the elastic lamina. Magenta indicates endothelial cells. Images obtained using 25× magnification with exposure time of 400 msec. TUNEL staining for apoptosis revealed no evidence of endothelial cell apoptosis in the uninjured control carotid arteries, with a prominent endothelial cell monolayer (white arrowhead). Mild apoptosis was noted in the media of balloon‐injured left carotid arteries, as expected. Black arrows indicate apoptotic cells. Representative images obtained using 40× magnification. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28242820), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: CD31/PECAM-1 Antibody [NB100-2284] - The growth of a patient mesothelioma xenograft was supported by development of a murine vascular network. A patient mesothelioma was implanted in a cohort of mice & monitored for tumor growth. Once tumors began to actively grow, representative tumors were resected at weekly intervals & analyzed for vessel content. Staining for huCD31 was prominent in the original patient specimen (A), much reduced at 4 weeks (B) & negligible at 9 weeks (C). In contrast, patient specimens were not stained for msCD34 (D), whereas at 4 weeks large numbers of vessels stained for msCD34 (E) & by 9 weeks, msCD34 labeled vessels were predominant (F). The graph (G) summarizes the loss of detectable human vessels & acquisition of murine vessels over a 9 week period. (bars = 100 μ). Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: CD31/PECAM-1 Antibody [NB100-2284] - TGF‐ beta 1 released by MSCs contributes to induced myofibroblast differentiation & granulation tissue formationA–C2.5 × 105 of TGF‐ beta 1 siRNA or control siRNA‐transfected AT‐MSCs were intradermally injected around each of CD18−/− murine wound. PBS mock injection served as negative control. Wound tissue was harvested at day 2, 5, & 7 post‐wounding for quantification of human TGF‐ beta 1 mRNA (A) at day 2 by qPCR, total TGF‐ beta 1 (B), & active TGF‐ beta 1 (C) protein at day 5 by ELISA. Data are expressed as mean ± SEM, n = 3 wounds per group, **P < 0.01, by one‐way ANOVA with Tukey's test.D–GExpression of alpha ‐SMA (D & E) & CD31 (F & G) at days 5 & 7 by immunostaining on tissue sections. The dashed lines indicate the border of the wound bed & epidermis or eschar. e, epidermis; es, eschar; wb, wound bed. Scale bars: 100 μm. Quantification data are expressed as mean ± SEM, n = 3 wounds per group, *P < 0.05, ***P < 0.001, by one‐way ANOVA with Tukey's test.Source data are available online for this figure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32080965), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: Rabbit Polyclonal CD31/PECAM-1 Antibody [NB100-2284] - CD31 positive vessels in a canine splenic hemangiosarcoma.Tissue slides were loaded into automated research stainer, dewaxed and pretreated with EDTA-based epitope retrieval ER2 solution for 20 mins at 100°C. After a peroxide incubation for 5 min, the rabbit antibody against CD31 (NB100-2284) (1:100) was incubated for 60 mins at room temperature followed by Bond Polymer (anti-rabbit HRP) incubation for 8 min at room temperature. Mixed DAB reagent (Polymer Refine Detection Kit) was incubated for 10 mins, and Hematoxylin (Refine Detection Kit) counterstaining for 10 mins. After staining, sample slides were washed in water, dehydrated using ethanol gradient (70%, 90%, 100%), washed three times, and mounted in mounting medium. Image from a verified customer review.

Reactivity:

Hu, Mu, Rt, Po, Ca

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Ca

Applications:

, ,

Applications: , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

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NB100-137

EGLN1/PHD2 Antibody - BSA Free

(12 Reviews)

(96 Publications)

0.1 mg

NB100-137

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Analysis of PHD2 expression in human prostate cancer cell lines. Western blot image submitted by a verified customer review.

Immunocytochemistry/Immunofluorescence: EGLN1/PHD2 Antibody [NB100-137] - Staining of endogenous PHD2 in U2OS cells. ICC/IF image submitted by a verified customer review.

<b>Genetic Strategies Validation. </b>Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Detection of PHD2 in human glioblastoma cell lysates. Image submitted by a verified customer review.

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Detection of human PHD2 by Western blot. Recombinant epitope-tagged PHD1, PHD2 or PHD3 (10 ng/lane) or whole cell lysate from HeLa cells. EGLN1/PHD2 antibody used at 1 ug/mL. Detection by chemiluminescence.

Immunohistochemistry: EGLN1/PHD2 Antibody [NB100-137] - EGLN1/PHD2 and EGFR expression levels positively correlate in breast cancer. Processed tissue microarrays of breast cancer biopsies from 313 patients were stained with EGLN1/PHD2 and EGFR antibodies. Four representative immunohistochemistries of human breast cancer with low and high expression of EGLN1/PHD2 and EGFR are shown. Magnification 10x. Image collected and cropped by CiteAb from the following publication (//www.oncotarget.com/article/14241/text/) licensed under a CC-BY license.

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Exogenous PHD2 has no impact on the gene expression of HIF-1alpha and HIF-2alpha. U87MG cells were transfected with empty vector (pcDNA) or PHD2 plasmid. Twenty-four hours after transfection, PHD2 was detected by immunoblotting. beta-Actin was used as loading control. The results are representative for three independent experiments. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2014295), licensed under a CC-BY license.

Flow Cytometry: EGLN1/PHD2 Antibody [NB100-137] - An intracellular stain was performed on Jurkat cells with NB100-137AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Analysis of PHD2 in normoxia (lane 1) or hypoxia (lane 2) treated SK-N-BE(2) cells using anti-EGLN1/PHD2 antibody. Western blot image submitted by a verified customer review.

Immunocytochemistry/Immunofluorescence: EGLN1/PHD2 Antibody [NB100-137] - Staining of PHD2 in HeLa cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).

Immunohistochemistry: EGLN1/PHD2 Antibody [NB100-137] - Staining of lung vascular endothelium with EGLN1/PHD2 antibody. Image at 40X.

Flow Cytometry: EGLN1/PHD2 Antibody [NB100-137] - Detection of PHD2 in Jurkat cells. One million Jurkat cells were fixed, permeabilized, and stained with 3.0 ug/mL anti-EGLN1/PHD2 antibody in a 150 uL reaction. Isotype control (black), anti-MLL1 (red).

PHD2 and EGFR expression levels positively correlate in breast cancerProcessed tissue microarrays of breast cancer biopsies from 313 patients were stained with PHD2 and EGFR antibodies (cf Materials and methods). Four representative immunohistochemistries of human breast cancer with low and high expression of PHD2 and EGFR are shown. Magnification 10x.

Initial characterization of the OS-9 protein.(A) OS-9 expression in various human cell lines. Equal protein amounts of total cell lysates were used for SDS-PAGE & subsequent Western blotting. For each cell line, two independent samples are shown. Endogenous OS-9 was detected with a polyclonal antibody raised against a peptide corresponding to amino acids 600–667 of isoform 1 of OS-9. (B) Protein stability assay of endogenous OS-9. U2OS cells were treated with the translational inhibitor cycloheximide (100 µM). At indicated time points, whole cell lysates were analysed by immunoblotting. (C) Effect of hypoxia on OS-9 expression. For hypoxia, UT-7 cells were exposed to 1% O2 for 24 h prior to Western blot analysis. To determine any influence of HIF-1 alpha on OS-9 expression under normoxia, cells were incubated with the prolyl hydroxylase inhibitor DMOG (0.5 mM) for 24 h. (D) Protein interaction between OS-9 & PHD2 in vitro. For co-immunoprecipitation, U2OS cells were transiently co-transfected with the plasmids pOS-9-V5 & pPHD2-His, lysed in NP40 buffer, & subjected to immunoisolation with anti-V5 antibody recognizing OS-9 by its V5-tag. OS-9 & its associated proteins were separated by SDS-PAGE & analyzed by Western blot (lane 2). As controls, samples of untransfected (lane 1) cells or cells transfected with a single plasmid (lanes 3–4) were loaded. Representative Western blots are shown for each subfigure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/21559462), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Cellular localization of OS-9 & PHD2.(A) A lectin gel-shift assay was conducted to test for glycosylated proteins. Total cell lysates of U2OS cells were incubated in the presence or absence of the endoglycosidases EndoH & PNGaseF for 6 h at 37°C. Digest products were separated on a reducing 10% SDS-PAGE gel which contained concanavalin A co-polymerized in the top layer of the separating gel to retard mobility of glycosylated proteins [56]. Glycosylated OS-9 is indicated as ‘g’, deglycosylated OS-9 as ‘d’. (B) Detection of OS-9 & PHD2 in the nuclear fraction. HEK293 cells with & without transfection of the plasmid pcDNA3-OS-9 were separated into nuclear fraction (N) & postnuclear supernatant (PS), the latter containing cytoplasm & organelles. Western blot analysis included BiP, GAPDH & lamin A as typical marker proteins for the ER, the cytoplasm & the nucleus, respectively. (C) Detection of OS-9 & PHD2 in the cytoplasm. HEK293 cells were co-transfected with pcDNA3-OS-9 & pPHD2-V5. For hypoxia, cells were exposed to 3% O2 for 4 h. Cells were treated with 50 µg/ml digitonin & centrifuged to obtain a cytoplasmic (C) & an organelle fraction (O) & subjected to immunoblotting. (D) Isolation of cellular endomembranes. HEK293 cells were lysed mechanically by several passages through a 30½G needle. The postnuclear supernatant was processed further by ultra-centrifugation to separate the organelles (O) from the cytosol (C). High salt treatment (1 M KCl) of the organelle fraction produced a wash fraction (W) that contained dissociated peripheral membrane proteins. For immunoblot analysis of subcellular fractionations, cell aliquots were normalized for cell number prior to loading (B–D). Representative Western blots are shown for each subfigure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/21559462), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Representative picture of western blot in histopathologically unchanged tissue (N) & primary cancerous tissue (C) from patients with CRC. Immunodetection of bands was performed with Rp anti- PHD1, - PHD2, - PHD3 & - FIH Ab, followed by incubation with goat anti-rabbit HRP-conjugated Ab. The membrane was stripped & 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) & Biospectrum® Imaging System 500, UVP Ltd. (Upland, CA). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24195777), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - OS-9 shows no effect on regulation of HIF-1 alpha .Total cell lysates were used for SDS-PAGE & subsequent Western blotting. To generate nearly anoxic conditions, cells were exposed to an oxygen consuming chemical system or to 1% or 3% O2 for 4 h to generate hypoxia. (A) U2OS, HeLa & Hep3B cells were transiently transfected with the plasmid pOS-9-V5 48 h prior to the experiment. Lamin A & actin were used as loading controls. (B) U2OS cells were subjected to ER stress by incubation either with tunicamycin (1 µg/ml) or thapsigargin (0.5 µg/ml) for 20 h. To detect HIF-1 alpha under normoxia, cells were treated with DMOG (1 mM) for 4 h. A sample of DMSO-only treated cells was loaded to exclude unspecific side effects of the solvent. (C) U2OS cells were transduced with lentiviral construct pLKO.1-shRNA-OS-9 (shOS-9) mediating a stable knockdown of OS-9 expression. Control cells (c) were transduced with plasmid pLKO.1-puro. Representative Western blots are shown for each subfigure. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/21559462), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Oxygen chemotaxis is independent of the PHD/HIF pathway. a–dHIF1A, HIF2A, HIF1A+HIF2A & PHD2 CRISPR/Cas9 KO clones characterisation, respectively, regarding O2-directed migration. Left panels: immunoblot validation of HIF1A, HIF2A, HIF1A+HIF2A & PHD2 KO clones. To blot HIFs factors (a, b, c), cells were first pre-treated for 5 h with CoCl2 300 µM before protein extraction, a condition that promotes HIF factors accumulation (cf. Supplementary Fig. 8f). Middle panels: cell trajectories under confinement. Red dashed lines indicate the border of the cell cluster at 0 h. Right panels: relative distribution of MCF10A KO clones versus wt cells at the edge of the cluster at 48 h. These experiments demonstrate that HIF factors deletion does not prevent aerotaxis. e, f Tracks & redistribution of wt & PHD2 KO clone silenced for PHD3 (siPHD3) or not (siCTR). g Tracks & redistribution of MCF10A cells treated with DMOG (50 µM) or CoCl2 (50 µM) to inhibit PHDs, or with vehicle only (DMSO). These experiments demonstrate that PHDs do not participate in O2-sensing during aerotaxis. Confinement was applied for 48 h (a–g). Scale bars, 500 µm Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30382089), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - PHD3 expression correlates with a mesenchymal-like morphology in pancreatic ductal adenocarcinoma cell lines.NHF-1 (Fibroblast) MiaPaca2, Panc1, CAPAN1 & BxPC3 cells were harvested for RNA & protein following 24 hours exposure to normoxia (21% O2) or hypoxia (1% O2). (A) Phase-contrast images at 10× magnification were taken of MiaPaca2 (mesenchymal-like) morphology & BxPC3 cells (differentiated, epithelial morphology) under normoxic conditions. (B) PHD3 mRNA expression was determined by qRT-PCR & graphed relative to BxPC3 in normoxia. All samples were normalized to 18S rRNA & graphed as expression relative to BxPC3-Vec Normoxia (lane 1). n = 3, error bars = 1 S.D. (C) Whole cell lysate was resolved by SDS-PAGE & blotted for beta -tubulin PHD3 & PHD2. N = normoxia, H = hypoxia (1% O2). Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0083021), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - Iron supplementation restores HIF-1 alpha levels to normal following ATP6V0E1 inhibition in HeLa cells. (A) Schematic diagram of the multimeric V-ATPase complex. (B) Chemical inhibition of V-ATPase by 10 nM BafA treatment for 24 h, increased HIF-1 alpha levels in HIF alpha -GFPODD reporter cells. Treatment with 100 μM Fe (III) citrate significantly reduced the elevated HIF-1 alpha levels associated with loss of ATP6V0E1 (1 × 106 cells per sample harvested & analyzed; N = 2). (C) Knock-down of ATP6V0E1 subunit with three different CRISPR-Cas9 guides resulted in significant upregulation of HIF-1 alpha levels in HIF alpha -GFPODD reporter cells. Co-treating cells with 100 μM Fe (III) citrate led to a reduction in HIF-1 alpha levels across the three ATP6V0E1 depleted cells. phd2 was knocked down as a control & treatment with 100 μM Fe (III) citrate did not result in reduction of HIF-1 alpha levels (1 × 106 cells per sample harvested & analyzed; N = 2). FACs plot shown is a representative image of two biological repeats performed. (D) Immunoblot analysis for HIF-1 alpha & PHD2 levels in HIF alpha -GFPODD reporter cells with either ATP6V0E1 or PHD2 depleted or treated with 10 nM BafA. The cells were treated with 100 μM Fe (III) citrate for 24 h. beta actin was used as a control. Results validated findings observed by flow cytometry, whereby HIF-1 alpha levels were upregulated following ATP6V0E1 knock-down or inhibition & levels were re-normalized upon Fe (III) citrate treatment. Treatment of Fe (III) citrate in PHD2 depleted cells did not alter HIF-1 alpha levels. All experiments were performed in biological duplicate. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32984302), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: EGLN1/PHD2 Antibody [NB100-137] - PHD3 depletion stabilizes hypoxic p27 expression by increasing p27 half-life. a Cell cycle arrest at G0 & 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 & 786-O cells. Asterisk indicates significant difference (p < 0,05; n = 3). c Cell cycle arrest at G0 & 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 & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26223520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Pm, Pm-Or, Rb, RM

Species Glossary

Reactivity: Hu, Mu, Rt, Pm, Pm-Or, Rb, RM

Applications:

, , , , , , ,

Applications: , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-137AF350
Alexa Fluor 405	NB100-137AF405
Alexa Fluor 488	NB100-137AF488
Alexa Fluor 532	NB100-137AF532
Alexa Fluor 594	NB100-137AF594
Alexa Fluor 647	NB100-137AF647
Alexa Fluor 700	NB100-137AF700
Alexa Fluor 750	NB100-137AF750
Allophycocyanin	NB100-137APC
Biotin	NB100-137B
CoraFluor 1	NB100-137CL1
DyLight 350	NB100-137UV
DyLight 405	NB100-137V
DyLight 488	NB100-137G
DyLight 550	NB100-137R
DyLight 594	NB100-137DL594
DyLight 650	NB100-137C
DyLight 680	NB100-137FR
DyLight 755	NB100-137IR
FITC	NB100-137F
HRP	NB100-137H
Janelia Fluor 525	NB100-137JF525
Janelia Fluor 549	NB100-137JF549
Janelia Fluor 646	NB100-137JF646
Janelia Fluor 669	NB100-137JF669
PE	NB100-137PE
PerCP	NB100-137PCP
mFluor Violet 450 SE	NB100-137MFV450
mFluor Violet 500 SE	NB100-137MFV500
mFluor Violet 610 SE	NB100-137MFV610

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NB100-131

HIF-1 alpha Antibody (ESEE122) - BSA Free

(3 Reviews)

(89 Publications)

0.1 ml

NB100-131

Simple Western: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Image shows a specific band for HIF-1 alpha in 0.5 mg/mL of Hypoxic HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Detection of HIF-1 alpha (red dye 568) in a cultured raw mouse macrophage cell line, using NB100-131. Photos courtesy of Susan Alexander and Hattie Gresham, PhD.

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Detection of HIF-1 alpha (red dye) in a cell cytospin from a lavage of a murine skin pouch infected with S. aureus. 100X magnification. Blue: DAPI nuclear staining.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Analysis of a FFPE mouse kidney tissue section using HIF-1 alpha antibody clone ESEE122 at 1ug/mL concentration. The detection was performed using X-cell plus universal HRP polymer detection system with Vector SG chromagen substrate. Image courtesy of a product review by Steven Grover.

Immunohistochemistry: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Immunohistochemical staining of HIF-1 alpha in normal renal tissue (A) and clear cell renal cell carcinoma (CCRCC) (D). A homogeneous cytoplasmic staining of tubular cells and weak staining in glomerules was observed with HIF-1 alpha (A). 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 x200). Image collected and cropped by CiteAb from the following publication (//www.jeccr.com/content/28/1/40), licensed under a CC-BY license.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Analysis of HIF-1 alpha in paraffin-embedded mouse kidney tissue section using anti-HIF-1 alpha antibody. Image from verified customer review.

<b>Biological Strategies Validation. </b>Immunohistochemistry: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Nuclear HIF-1 alpha protein expression. Xenografts were established in female nude mice (BALB c[nu/nu]) by subcutaneous injection of head and neck squamous cell carcinoma cell lines UT-SCC-2 (UT-2) and UT-SCC-14 (UT-14). Cetuximab (1 mg/injection) or PBS was administered by intraperitoneal injection at day 10, 14, and 17. A tissue microarray was constructed from tumours harvested at day 21, and the expression of nuclear HIF-1 alpha was evaluated by immunohistochemistry (IHC) in untreated controls and cetuximab-treated tumour specimens. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28756482/) licensed under a CC-BY license.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Negative control stain of human placenta (from sea level) using mouse IgG at 1:100. 4uM paraffin-embedded section.

$Immunohistochemistry-Paraffin: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Analysis of a FFPE tissue section of human renal cancer xenograft using HIF-1 alpha antibody (NB100-131 Lot 83115) at 1:200 dilution. The antibody generated a strong cytoplasmic staining mainly in the cancer cells. Only a fraction of cells depicted nuclear staining, while weak to negligible positivity was seen in the tumor stromal cells.$

Immunohistochemistry: HIF-1 alpha Antibody (ESEE122) [NB100-131] - HIF-1 alpha staining in hypoxia-induced human placenta.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Representative immunohistochemical expression for HIF-1alpha, c-Met, CA9 and GLUT1. HIF-1alpha is stained in cytoplasm shown with no staining in normal cervix (A), weak staining intensity in high grade CIN (B), and strong staining intensity in squamous cell carcinoma (C). Scale bar: 50 um. Image collected and cropped by CiteAb from the following publication (//www.translational-medicine.com/content/11/1/185), licensed under a CC-BY license.

Positive immunohistochemical staining for (A) VEGF, (B) HIF-1 alpha , (C) Dll4 (tumor cells), (D) Dll4 (endothelial cells), and (E) CD31 (for microvessel counting, ×200 magnification).

Immunohistochemical analysis in non-GIST STS representing negative, and score 1–3 of CAIX, GLUT-1, HIF-1 alpha , and HIF-2 alpha . non-GIST STS: non-gastrointestinal stromal tumor soft-tissue sarcomas, CAIX: carbonic anhydrase IX, GLUT-1: glucose transporter-1, and HIF-1/2 alpha : hypoxia induced factor 1/2 alpha .

Immunohistochemistry: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Immunohistochemical analysis in non-GIST STS representing negative, & score 1–3 of CAIX, GLUT-1, HIF-1 alpha , & HIF-2 alpha . non-GIST STS: non-gastrointestinal stromal tumor soft-tissue sarcomas, CAIX: carbonic anhydrase IX, GLUT-1: glucose transporter-1, & HIF-1/2 alpha : hypoxia induced factor 1/2 alpha . Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/22454562), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Digital image analysis of cytoplasmic & membranous staining. Cytoplasmic HIF-1 alpha staining is shown (A) & automated image analysis utilizing TissueIA recognizes cytoplasmic HIF-1 alpha staining highlighted in green color (B). CA9 is shown in membranous staining (C) & automated image analysis determines membranous CA9 staining highlighted in green color (D). The output from the algorithm returns a number of quantitative measurements for intensity & percentage of positive staining present. Scale bar: 100 μm. Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-185), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Immunohistochemical staining of HIF-1 alpha , VEGF-A & VEGF-C in normal renal tissue (A-C) & clear cell renal cell carcinoma (CCRCC) (D-F). A homogeneous cytoplasmic staining of tubular cells & weak staining in glomerules was observed with HIF-1 alpha (A), while VEGF-A & VEGF-C were positive in tubular cells, glomerular mesangium & interstitial macrophages (B & C). In CCRCC, HIF-1 alpha immmunoreactivity was nuclear and/or cytoplasmic (D), while it was perimembranous and/or diffuse cytoplasmic for VEGF-A & VEFG-C (E & F). (magnification ×200). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/19302703), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Positive immunohistochemical staining for (A) VEGF, (B) HIF-1 alpha , (C) Dll4 (tumor cells), (D) Dll4 (endothelial cells), & (E) CD31 (for microvessel counting, ×200 magnification). Image collected & cropped by CiteAb from the following publication (//www.spandidos-publications.com/10.3892/or.2012.2075), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: HIF-1 alpha Antibody (ESEE122) [NB100-131] - Representative immunohistochemical expression for HIF-1 alpha , c-Met, CA9 & GLUT1. HIF-1 alpha is stained in cytoplasm shown with no staining in normal cervix (A), weak staining intensity in high grade CIN (B), & strong staining intensity in squamous cell carcinoma (C). c-Met (D-F), CA9 (G, H) & GLUT1 (I) shows cell membranous staining. Representative c-Met expression in cervical samples shown with no staining in normal cervix (D), weak membranous staining intensity in squamous cell carcinoma (E) & strong intensity in squamous cell carcinoma (F). CA9 expression showing moderate intensity staining in carcinoma in situ (CIS) (G) & strong staining in adenocarcinoma (H). GLUT1 expression showing strong intensity in squamous cell carcinoma (I). Scale bar: 50 μm. Image collected & cropped by CiteAb from the following publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-185), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

CASQ2 affects the TME and invasiveness through HIF1 alpha . (A) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis using the mouse genome (Differentially expressed genes, DEGs; n = 530). (B) The heatmap for average of expression (FPKM) value of most significant differentially expressed genes of angiogenesis and the HIF1 alpha signaling pathway. (C) The mRNA level of Hif1a, Pgf, Flt1, Kdr, Flt4, and Nos3 in Hs578T‐CTL and CASQ2 o/e tumors (means +/- SEM, n = 3; ***P < 0.001 by the multiple t‐test). (D, E) Expression level of alpha ‐SMA, FSP, and HIF1 alpha in WI‐38 cells after indirect coculture with Hs578T‐CTL or Hs578T‐CASQ2 o/e cells (means +/- SEM, n = 3; *P < 0.05 by the multiple t‐test). (F, G) Expression level of CASQ2 and vimentin in Hs578T‐CTL or Hs578T‐CASQ2 o/e cells indirectly cocultured with WI‐38 (means +/- SEM, n = 3; *P < 0.05 by the multiple t‐test). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34743414), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

CASQ2 affects the tumor invasiveness through HIF1 alpha . (A, B) Expression level of HIF1 alpha in caffeine‐treated Hs578T cells with or without overexpression of CASQ2 (means +/- SEM, n = 3; *P < 0.05, **P < 0.01 by the Mann–Whitney U‐test). (C) Confocal microscopy images showing F‐actin in Hs578T cells overexpressing CASQ2 under hypoxic conditions (**P < 0.01 by the Mann–Whitney U‐test). Scale bar = 10 μm. (D) Proliferation rate of control (CTL) and CASQ2‐overexpressing (CASQ2 o/e) breast cancer cells under normoxic and hypoxic conditions (mean +/- SEM, n = 3; *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 by ANOVA with Tukey’s post hoc test). (E, F) Hematoxylin and eosin staining of lung tissue sections from Hs578T‐CTL‐ or CASQ2‐derived tumors (mean +/- SEM, n = 5; *P < 0.05 and **P < 0.01 by two‐tailed Student’s t‐test). Scale bar = 100 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34743414), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

CASQ2 induces phenotypic changes in breast cancer cells. (A) Relationship between the proliferation rate and overexpression of CASQ2 in breast cancer cell lines (mean +/- SEM, n = 3; *P < 0.05, **P < 0.01, and ***P < 0.001 using the multiple t‐test). (B) Migration and invasion rates of Hs578T cells (mean +/- SEM, n = 3; **P < 0.01 using two‐tailed Student’s t‐test). Scale bar = 50 μm. (C) Three‐dimensional culture of Hs578T cells (mean +/- SEM, n = 3; **P < 0.01 using two‐tailed Student’s t‐test). Scale bar = 100 μm. (D) Tumorsphere culture of Hs578T cells (mean +/- SEM, n = 3; **P < 0.01 using two‐tailed Student’s t‐test). Scale bar = 100 μm. (E) Expression of CD44, CD24, and ALDH1 cancer stem cell markers in adherent and tumorsphere cultures of Hs578T cells. (F) Measurement of intracellular Ca2+ in Hs578T cells using a calcium crimson reagent. Cells were loaded with the calcium indicator calcium crimson (5 μm) for 30 min, and then, BAPTA‐AM (10 μm) was added. After washing, the cells were stimulated by treatment with 5 μm caffeine to measure the intracellular calcium concentration at 360 s (mean +/- SEM, n = 3; *P < 0.05 and **P < 0.01 after ANOVA with Tukey’s post hoc test). (G) Effect of lacidipine on the expression of cancer stem cell markers in tumorspheres of breast cancer cells. The figure shows one of three independent experiments. Statistical test result is shown in Fig. S6. (H) Epithelial–mesenchymal transition (EMT)‐related protein expression in breast cancer cells. All results are representatives of three independent experiments. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34743414), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Bv, Ca

Species Glossary

Reactivity: Hu, Mu, Rt, Bv, Ca

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-131AF350
Alexa Fluor 405	NB100-131AF405
Alexa Fluor 488	NB100-131AF488
Alexa Fluor 532	NB100-131AF532
Alexa Fluor 594	NB100-131AF594
Alexa Fluor 647	NB100-131AF647
Alexa Fluor 700	NB100-131AF700
Alexa Fluor 750	NB100-131AF750
Allophycocyanin	NB100-131APC
Biotin	NB100-131B
CoraFluor 1	NB100-131CL1
DyLight 350	NB100-131UV
DyLight 405	NB100-131V
DyLight 488	NB100-131G
DyLight 550	NB100-131R
DyLight 594	NB100-131DL594
DyLight 650	NB100-131C
DyLight 680	NB100-131FR
DyLight 755	NB100-131IR
FITC	NB100-131F
HRP	NB100-131H
Janelia Fluor 525	NB100-131JF525
Janelia Fluor 549	NB100-131JF549
Janelia Fluor 646	NB100-131JF646
Janelia Fluor 669	NB100-131JF669
PE	NB100-131PE
PerCP	NB100-131PCP
mFluor Violet 450 SE	NB100-131MFV450
mFluor Violet 500 SE	NB100-131MFV500
mFluor Violet 610 SE	NB100-131MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-131SS
Azide and BSA Free	NBP2-80762

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NB600-562

CD31/PECAM-1 Antibody (JC/70A) - BSA Free

(4 Reviews)

(54 Publications)

0.1 ml

NB600-562

CD31/PECAM-1 was detected in immersion fixed THP-1 (red, positive) but not THP-1 KO (negative) cells using 8 ug/mL Mouse Anti-Human CD31/PECAM-1 (JC/70A) Monoclonal Antibody (Catalog # NB600-562) for 3 hours at room temperature. Cells were stained with the NorthernLights(TM) 557-conjugated Donkey anti-Mouse IgG Secondary Antibody (red, Catalog # NL007)) and and counterstained with DAPI (blue).

<b>Genetic Strategies Validation and Orthogonal Strategies Validation.</b>Formalin-fixed paraffin-embedded tissue sections of human metastatic tonsil were probed for CD31 mRNA (ACD RNAScope probe, catalog # 487381; Fast Red chromogen, ACD catalog # 322500). Adjacent tissue section was processes for immunohistochemistry using mouse monoclonal (NB600-562) at 1:25 dilution for 1 hour at room temperature followed by incubation with the anti-mouse IgG VisUCyte HRP Polymer Antibody (Catalog # VC001) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue).

Rabbit Heart, CD31 Stained Red. Image from verified customer review.

A cell surface stain was performed on HUVEC cells with ( NB600-562, blue) along with a matched isotype control NBP2-27287 (orange). Cells were incubated in an antibody dilution of 1:100 for 20 minutes at RT. (Panel A). A negative control (HeLa cells) was also stained to ensure antibody specificity (Panel B).

GMF-beta is involved in neovasculogenesis in human glioblastoma. A. Co-expression of GMF-beta and CD31/PECAM-1 in several tumor cells. B. The GMF-beta+/CD31+ incomplete microvessel-like structure in hypovascular zones. C. GMF-beta+/CD31+ immature microvessel in hypovascular zones. D. Single GMF-beta+/CD31+ mature microvessel in vascularized areas. E. GMF-beta-/CD31+ mature microvessels in vascularized areas. Red arrows denote GMF-beta staining, black arrows indicate CD31/PECAM-1 staining. Scale bar: 50 um. Image collected and cropped by CiteAb from the following publication (//www.oncotarget.com/abstract/5509), licensed under a CC-BY license.

Analysis of CD31/PECAM1 in human spleen using DAB with hematoxylin counterstain.

Analysis of CD31/PECAM1 expression in 1) Jurkat whole cell lysate, 2) human platelet lysate and 3) U937 whole cell lysate.

Comparison of the expression of NMDAR1 on vessels and EndoMT markers in the urinary bladder.Biopsy specimens of frozen sections from the urinary bladders of (A, C, E) normal control subjects and (B, D, F) KC patients. (A, B) Double-immunofluorescence staining with antibody to endothelial marker CD31 (green) and NMDAR1 (red); DAPI was used as a nuclear stain (blue); co-expression of CD31 and NMDAR1 was evident in two groups (yellow, denoted by white arrows). (CF) Double-immunofluorescence staining with antibody to endothelial marker CD31 (green) and two mesenchymal markers: FSP1 (red, in panels C and D) and -SMA (red, in panels E and F); DAPI was used as a nuclear stain (blue); co-expression of CD31 and FSP1 or -SMA was evident in three groups (yellow, denoted by white arrows) (white bars: 50 m). Microvascular Injury in Ketamine-Induced Bladder Dysfunction. PLoS One (2016)

A surface stain was performed on Jurkat cells with the Dylight 550 conjugate of CD31/PECAM-1 Antibody (JC/70A) (NB600-562R, blue) and a matched isotype control (orange). Cells were incubated in an antibody dilution of 5 ug/mL for 20 minutes at room temperature. Both antibodies were conjugated to DyLight 550.

HUVEC 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 CD31/PECAM-1 Antibody (JC/70A) at 2 ug/ml overnight at 4C and detected with an anti-mouse 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.

Imaging of Feline Endothelial cells with antibody dilution of 1:25. CD31/PECAM-1 (Red), DAPI (Blue). This image was submitted via customer Review.

FFPE human tonsil stained with CD31/PECAM-1 Antibody (JC/70A).

A cell surface stain was performed on Jurkat cells with the Alexa Fluor 488 conjugate of CD31/PECAM-1 Antibody (JC/70A) (NB600-562AF488, blue) along with a matched isotype control. Cells were incubated in an antibody dilution of 10 ug/ml for 20 minutes at RT. Both antibodies were conjugated to Alexa Fluor 488.

A cell surface stain was performed on THP-1 cells with the Alexa Fluor 488 conjugate of CD31/PECAM-1 Antibody (JC/70A) NB600-562AF488 (blue) along with a matched isotype control. Cells were incubated in an antibody dilution of 5 ug/ml for 20 minutes at RT. Both antibodies were conjugated to Alexa Fluor 488.

A surface stain was performed on THP-1 cells with the Alexa Fluor 647 conjugate of CD31/PECAM-1 Antibody (JC/70A) (NB600-562AF647, blue) and a matched isotype control (orange). Cells were incubated in an antibody dilution of 2.5 ug/mL for 20 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Immunocytochemistry/ Immunofluorescence: CD31/PECAM-1 Antibody (JC/70A) - BSA Free [NB600-562] - Immunohistochemial comparison of the expression of NMDAR1 on vessels & EndoMT markers in the urinary bladder.Biopsy specimens of frozen sections from the urinary bladders of (A, C, E) normal control subjects & (B, D, F) KC patients. (A, B) Double-immunofluorescence staining with antibody to endothelial marker CD31 (green) & NMDAR1 (red); DAPI was used as a nuclear stain (blue); co-expression of CD31 & NMDAR1 was evident in two groups (yellow, denoted by white arrows). (C–F) Double-immunofluorescence staining with antibody to endothelial marker CD31 (green) & two mesenchymal markers: FSP1 (red, in panels C & D) & alpha -SMA (red, in panels E & F); DAPI was used as a nuclear stain (blue); co-expression of CD31 & FSP1 or alpha -SMA was evident in three groups (yellow, denoted by white arrows) (white bars: 50 μm). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27529746), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

IF staining of IPAH patient CD31/PECAM-1 lung slice. Image from a verified customer review.

Reactivity:

Hu, Mu, Fe, Rb

Species Glossary

Reactivity: Hu, Mu, Fe, Rb

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB600-562AF350
Alexa Fluor 405	NB600-562AF405
Alexa Fluor 488	NB600-562AF488
Alexa Fluor 532	NB600-562AF532
Alexa Fluor 594	NB600-562AF594
Alexa Fluor 647	NB600-562AF647
Alexa Fluor 700	NB600-562AF700
Alexa Fluor 750	NB600-562AF750
Allophycocyanin	NB600-562APC
Biotin	NB600-562B
CoraFluor 1	NB600-562CL1
DyLight 350	NB600-562UV
DyLight 405	NB600-562V
DyLight 488	NB600-562G
DyLight 550	NB600-562R
DyLight 594	NB600-562DL594
DyLight 650	NB600-562C
DyLight 680	NB600-562FR
DyLight 755	NB600-562IR
FITC	NB600-562F
HRP	NB600-562H
Janelia Fluor 525	NB600-562JF525
Janelia Fluor 549	NB600-562JF549
Janelia Fluor 646	NB600-562JF646
Janelia Fluor 669	NB600-562JF669
PE	NB600-562PE
PerCP	NB600-562PCP
mFluor Violet 450 SE	NB600-562MFV450
mFluor Violet 500 SE	NB600-562MFV500
mFluor Violet 610 SE	NB600-562MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB600-562SS
Azide and BSA Free	NBP2-80641

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NB100-64796

CD31/PECAM-1 Antibody (TLD-3A12) - BSA Free

(1 Review)

(61 Publications)

0.25 mg

NB100-64796

Immunohistochemistry-Paraffin: CD31/PECAM-1 Antibody (TLD-3A12) [NB100-64796] - Analysis of CD31 in 4th day cutaneous wound section of diabetic rat using CD31/PECAM-1 Antibody (TLD-3A12). Image from verified customer review.

Immunohistochemistry-Paraffin: CD31/PECAM-1 Antibody (TLD-3A12) [NB100-64796] - IHC analysis for the expression of CD31 of wound tissues of different groups of rats (magnification 200). n = 6, **P less than 0.01. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/sci/2019/9564869/) licensed under a CC-BY license.

(a) HE staining for wound tissues of different groups of rats (magnification ×40). (b) IHC analysis for the expression of CD31 of wound tissues of different groups of rats (magnification ×200). n = 6, ∗∗P < 0.01.

Reactivity:

Rt

Species Glossary

Reactivity: Rt

Applications:

, , , ,

Applications: , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-64796AF350
Alexa Fluor 405	NB100-64796AF405
Alexa Fluor 488	NB100-64796AF488
Alexa Fluor 532	NB100-64796AF532
Alexa Fluor 594	NB100-64796AF594
Alexa Fluor 647	NB100-64796AF647
Alexa Fluor 700	NB100-64796AF700
Alexa Fluor 750	NB100-64796AF750
Allophycocyanin	NB100-64796APC
Biotin	NB100-63701
Biotin	NB100-64796B
CoraFluor 1	NB100-64796CL1
DyLight 350	NB100-64796UV
DyLight 405	NB100-64796V
DyLight 488	NB100-64796G
DyLight 550	NB100-64796R
DyLight 594	NB100-64796DL594
DyLight 650	NB100-64796C
DyLight 680	NB100-64796FR
DyLight 755	NB100-64796IR
FITC	NB100-63703-100ug
FITC	NB100-64796F
HRP	NB100-64796H
Janelia Fluor 525	NB100-64796JF525
Janelia Fluor 549	NB100-64796JF549
Janelia Fluor 646	NB100-64796JF646
Janelia Fluor 669	NB100-64796JF669
PE	NB100-64796PE
PerCP	NB100-64796PCP
mFluor Violet 450 SE	NB100-64796MFV450
mFluor Violet 500 SE	NB100-64796MFV500
mFluor Violet 610 SE	NB100-64796MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-64796SS

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NB100-2219

EGLN1/PHD2 Antibody - BSA Free

(3 Reviews)

(47 Publications)

0.1 ml

NB100-2219

Immunohistochemistry-Paraffin: EGLN1/PHD2 Antibody [NB100-2219] - Analysis of an FFPE mouse lung section using 1:200 dilution of EGLN1/PHD2 antibody. The staining was developed using HRP conjugated anti-rabbit secondary antibody and DAB reagent. The antibody generated a specific staining in the cytoplasm and nuclei of alveolar as well as bronchiolar epithelial cells. Cytoplasmic staining was observed in almost all cells while the nuclear positivity was seen in a subset of cells only.

Western Blot: EGLN1/PHD2 Antibody [NB100-2219] - Detection of EGLN1/PHD2 in mouse kidney lysate. ECL exposure, 20 seconds.

Immunocytochemistry/Immunofluorescence: EGLN1/PHD2 Antibody [NB100-2219] - EGLN1/PHD2 antibody at 1:500 in HeLa cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).

Immunocytochemistry/Immunofluorescence: EGLN1/PHD2 Antibody [NB100-2219] - Analysis of EGLN1/PHD2 in ARPE-19 cells using anti-PHD2 antibody. ICC/IF image submitted by a veriifed customer review.

Immunohistochemistry: EGLN1/PHD2 Antibody [NB100-2219] - Staining of renal tubular epithelium in mouse using NB100-2219 at 2.5 ug/mL.

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	NB100-2219AF350
Alexa Fluor 405	NB100-2219AF405
Alexa Fluor 488	NB100-2219AF488
Alexa Fluor 532	NB100-2219AF532
Alexa Fluor 594	NB100-2219AF594
Alexa Fluor 647	NB100-2219AF647
Alexa Fluor 700	NB100-2219AF700
Alexa Fluor 750	NB100-2219AF750
Allophycocyanin	NB100-2219APC
Biotin	NB100-2219B
CoraFluor 1	NB100-2219CL1
DyLight 350	NB100-2219UV
DyLight 405	NB100-2219V
DyLight 488	NB100-2219G
DyLight 550	NB100-2219R
DyLight 594	NB100-2219DL594
DyLight 650	NB100-2219C
DyLight 680	NB100-2219FR
DyLight 755	NB100-2219IR
FITC	NB100-2219F
HRP	NB100-2219H
Janelia Fluor 525	NB100-2219JF525
Janelia Fluor 549	NB100-2219JF549
Janelia Fluor 646	NB100-2219JF646
Janelia Fluor 669	NB100-2219JF669
PE	NB100-2219PE
PerCP	NB100-2219PCP
mFluor Violet 450 SE	NB100-2219MFV450
mFluor Violet 500 SE	NB100-2219MFV500
mFluor Violet 610 SE	NB100-2219MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-2219SS

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NB600-101

Calreticulin Antibody - BSA Free

(29 Publications)

0.1 ml

NB600-101

Immunocytochemistry/Immunofluorescence: Calreticulin Antibody [NB600-101] - Immunofluorescence staining of Calreticulin in HCT15 colon cancer cells using NB600-101. Secondary antibody was conjugated with Alexa Fluor 488. Photo courtesy of Dr. Birkenkamp Demtroeder, Arhus University Hospital.

Immunohistochemistry-Paraffin: Calreticulin Antibody [NB600-101] - Analysis of a FFPE tissue section of human thyroid gland using 1:50 dilution of Calreticulin antibody. The signal was developed using HRP-DAB based detection method which followed counterstaining of the nuclei with hematoxylin. This representative section shows a strong positivity of Calreticulin in the follicular epithelial cells, wherein the signal was found to be very intense in the perinuclear region of the cells which correlates well with Endoplasmic reticulum localization of this protein. The para-follicular cells, endothelial cells of blood vessels (not the RBCs though) and the loose connective tissue in the section showed a weak cytoplasmic staining. Some staining was observed in the follicles/colloids also which is potentially the secreted form of Calreticulin.

Immunocytochemistry/Immunofluorescence: Calreticulin Antibody [NB600-101] - The Calreticulin antibody NB600-101 was tested in HeLa cells at a 1:250 dilution against DyLight 488 (Green). Alpha-tubulin and nuclei were counterstained against DyLight 550 (Red) and DAPI (Blue), respectively.

Simple Western: Calreticulin Antibody [NB600-101] - Lane view shows a specific band for Calreticulin in 0.5 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Calreticulin was detected in immersion fixed U-2 OS human osteosarcoma cell line using Rabbit anti-Calreticulin Affinity Purified Polyclonal Antibody conjugated to Alexa Fluor® 647 (Catalog # NB600-101AF647) (light blue) at 10 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 40X objective.

$Characterization of purified ICVs. A549 cells were infected with the WSN virus and submitted to subcellular fractionation at 14 hpi. a Western-blot analysis of subcellular fractions of mock-infected (left panel) and IAV-infected (right panel) A549 cells, upon sucrose gradient ultracentrifugation. The same amount of total protein was analyzed for each fraction. TfR: transferrin receptor; PDI: protein disulfide isomerase; Cal: calreticulin; Clathrin HC: Clathrin Heavy Chain. For gel source data, Supplementary Fig. 17. b TEM analysis of an aliquot of fraction 60–40% from IAV-infected cells, upon fixation and negative staining. Black arrows: ICVs. White arrows: vRNPs. Scale bar: large panel 200 nm, small pannel 100 nm Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29123131), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Reactivity:

Hu, Mu, Rt, Bv, Ha, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, Bv, Ha, 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	NB600-101AF350
Alexa Fluor 405	NB600-101AF405
Alexa Fluor 488	NB600-101AF488
Alexa Fluor 532	NB600-101AF532
Alexa Fluor 594	NB600-101AF594
Alexa Fluor 647	NB600-101AF647
Alexa Fluor 700	NB600-101AF700
Alexa Fluor 750	NB600-101AF750
Allophycocyanin	NB600-101APC
Biotin	NB600-101B
CoraFluor 1	NB600-101CL1
DyLight 350	NB600-101UV
DyLight 405	NB600-101V
DyLight 488	NB600-101G
DyLight 550	NB600-101R
DyLight 594	NB600-101DL594
DyLight 650	NB600-101C
DyLight 680	NB600-101FR
DyLight 755	NB600-101IR
FITC	NB600-101F
HRP	NB600-101H
Janelia Fluor 525	NB600-101JF525
Janelia Fluor 549	NB600-101JF549
Janelia Fluor 646	NB600-101JF646
Janelia Fluor 669	NB600-101JF669
PE	NB600-101PE
PerCP	NB600-101PCP
mFluor Violet 450 SE	NB600-101MFV450
mFluor Violet 500 SE	NB600-101MFV500
mFluor Violet 610 SE	NB600-101MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB600-101SS

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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

Kit Type

Preservative

Protein Or Peptide

Research Areas

Has Sample Size

Promotional Item

Species Glossary

HIF-1 alpha Antibody (H1alpha67)

HIF-1 alpha Antibody - BSA Free

HIF-1 alpha Antibody

HIF-1 alpha Antibody - BSA Free

HIF-1 alpha Antibody (H1alpha67)

CD31/PECAM-1 Antibody

EGLN1/PHD2 Antibody - BSA Free

HIF-1 alpha Antibody (ESEE122) - BSA Free

CD31/PECAM-1 Antibody (JC/70A) - BSA Free

CD31/PECAM-1 Antibody (TLD-3A12) - BSA Free

EGLN1/PHD2 Antibody - BSA Free

Calreticulin Antibody - BSA Free