| Formulation | Catalog # | Availability | Price | |
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| Azide and BSA Free | NBP2-80760 | |||
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| BSA Free | NB100-122SS | |||
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| BSA Free | NB100-479SS | |||
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| Azide and BSA Free | NBP2-80766 | |||
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| Carrier Free | MAB1536-SP | |||
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| BSA Free | NB100-428SS | |||
| Formulation | Catalog # | Availability | Price | |
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| BSA Free | NBP2-75978SS | |||
| Azide and BSA Free | NBP2-80765 | |||
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| BSA Free | NBP1-47181SS | |||
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415525228.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-31020241612625.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415532090.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-31020241552521.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415525236.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415532060.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-31020241612610.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-31020241554522.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415523943.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415523937.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-31020241612694.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415523933.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415535163.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415541970.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415532043.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415535192.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415525216.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415532057.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415541951.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415525295.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-3102024165710.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415535122.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415535175.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-3102024155459.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-3102024168213.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-3102024165739.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-3102024155454.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-31020241683940.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-31020241555218.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415541924.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-3102024155458.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-31020241682192.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415541934.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-31020241682138.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-105_mouse-monoclonal-hif-1-alpha-antibody-h1alpha67-310202415532037.jpg)
![<b>Genetic Strategies Validation. </b>Reduction of HIF-2alpha levels leads to protection in UV-triggered apoptosis, but not for apoptosis caused by glucose and serum starvation in 786-O cells. Parental 786-O or those either stably expressing wild-type VHLp19 or stably infected with a control vector (pSuperRetro) or a pool of two HIF-2alpha shRNAs vectors [21] were grown to confluence and lysed. Cell alpha-tubulin.](http://images.novusbio.com/fullsize/HIF-2-alpha-EPAS1-Antibody-Knockdown-Validated-NB100-122-img0033.jpg)
![Western Blot: Rabbit Polyclonal HIF-2 alpha/EPAS1 Antibody [NB100-122] - Analysis of HIF-2 alpha/EPAS1 antibody on mouse liver nuclear extracts. Image from a verified customer review.](http://images.novusbio.com/fullsize/antibody/nb100-122_rabbit-polyclonal-hif-2-alpha-epas1-antibody-western-blot-222024141448..jpg)
![Immunocytochemistry/ Immunofluorescence: HIF-2 alpha/EPAS1 Antibody - BSA Free [NB100-122] - Muscle repair following eccentric contraction–induced injury is concomitant with dynamic alterations of HIF2A & HIF1A expression in SCs.(A–C) Representative images of EDL myofibers from injured muscles at various time points (n >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.](http://images.novusbio.com/fullsize/nb100-122_rabbit-polyclonal-hif-2-alpha-epas1-antibody-31020241535648.jpg)
![Immunocytochemistry/ Immunofluorescence: HIF-2 alpha/EPAS1 Antibody - BSA Free [NB100-122] - Genetic ablation of HIF2A in QSCs leads to transient activation, proliferation, & differentiation of SCs.(B) Representative images of myofibers from SC-HIF2AKO mice & control littermates (n >50 myofibers from 5 mice/group; 10 dpr). Immunofluorescence of Pax7 (red), HIF2A (green), MyoD (purple), & DAPI (blue) staining revealed HIF2A–MyoD+ & HIF2A+MyoD– SCs (arrowheads) in SC-HIF2AKO & control mice, respectively. Scale bar: 10 μm. Image collected & cropped by CiteAb from following publication (//www.jci.org/articles/view/96208), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-122_rabbit-polyclonal-hif-2-alpha-epas1-antibody-3102024153569.jpg)
![Immunocytochemistry/ Immunofluorescence: HIF-2 alpha/EPAS1 Antibody - BSA Free [NB100-122] - EpEX & EpCAM activate a STAT3-HIF2 alpha signal for EpEX/EpCAM-mediated iPSC formation.(A) iPSCs were infected with two different EpCAM shRNAs (two clones, #1 & #2). The protein expressions of EpCAM & HIF2 alpha were detected by Western blotting. (B) MEFs were stimulated by EpEX (1 μg/mL) at the indicated times. Nuclear-translocation was detected with a specific antibody against HIF2 alpha (n = 3). (C) Immunofluorescence staining was performed to detect subcellular localization of HIF2 alpha . Nuclei were stained with DAPI. Scale bar: 10 μm. (D) MEFs were treated with STAT3 inhibitor (WP1066, 10 μM), & then stimulated with EpEX for 30 min. The nuclear-translocation of HIF2 alpha was detected by Western blotting with anti-HIF2 alpha antibody (n = 3). (E) iPSC morphology was observed at day 20 after induction. Reprogramming of Oct4-GFP MEFs was induced by transfection of OSKM, OE + EpEX, & KE + EpEX with or without STAT3 inhibitor WP1066, or HIF2 alpha shRNA (n = 3). Scale bar: 50 μm. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep41852), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-122_rabbit-polyclonal-hif-2-alpha-epas1-antibody-310202415532070.jpg)
![Gel Super Shift Assays: HIF-2 alpha/EPAS1 Antibody - BSA Free [NB100-122] - Manipulation of DNA methylation at HRE sequences alters HIF binding. (A) The Hypoxia Response Element (HRE) contains a CpG site that can be methylated which prevents HIF1 & HIF2 binding in vitro. EMSA of in vitro–translated HIF1 & HIF2 binding to 32P-labelled unmethylated & methylated HRE probes. Competition with 250X molar excess of unlabelled/unmethylated HRE probe (lanes 3, 7,12, 16) or unlabelled control HRE-free probe (lanes 4, 8, 13, 17). HIF1 & HIF2 complexes bound to unmethylated HRE or methylated HRE supershifted with anti-HIF1 alpha (lanes 9 & 18) & anti-HIF2 alpha (lanes 5 & 14). (B) RCC4-VHL cells were grown in normoxia (NT) or treated with 25 µM decitabine (5aza) or grown in hypoxia (1%) for 48 hrs & with the addition of 25 µM decitabine (1% + 5aza). QPCR was performed using primers specific to VEGF, uPAR, TGF alpha , GUS, U1AsnRNP1. Samples were normalised to relative expression of the housekeeping gene, ACTIN. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-018-21524-5), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-122_rabbit-polyclonal-hif-2-alpha-epas1-antibody-310202415532046.jpg)
![Immunocytochemistry/ Immunofluorescence: HIF-2 alpha/EPAS1 Antibody - BSA Free [NB100-122] - 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.](http://images.novusbio.com/fullsize/nb100-122_rabbit-polyclonal-hif-2-alpha-epas1-antibody-31020241612668.jpg)
![Immunocytochemistry/ Immunofluorescence: HIF-2 alpha/EPAS1 Antibody - BSA Free [NB100-122] - HIF2 alpha stabilization results in exocrine cell atrophy & expansion of duct-like tubular structures. (A) Body weight (left panel), pancreas weight (middle panel) & body/pancreas weight ratio (right panel) in Pdx1-Cre;HIF2dPA & control mice at 2 & 8 weeks of age. Data are presented as mean ± SD. (B) HIF2 alpha accumulation in Pdx1-Cre;HIF2dPA analyzed by Western blot with anti-HA antibody. Two independent two-week-old control & mutant mice are shown. beta -actin protein was used for loading control. Full-length blots are presented in Supplementary Fig. 2. (D) Immunofluorescence analysis of HIF2 alpha in two-week-old control pancreata. Endogenous HIF2 alpha expression is observed in islets (marked by an white asterisk) but not in exocrine tissue. (D) Robust HIF2 alpha accumulation in the pancreas of two-week-old Pdx1-Cre;HIF2dPA mice. Hematoxylin/Eosin-stained pancreatic sections from P0 (E,F), two- (I,J) & eight-week-old (M,N) Pdx1-Cre;HIF2dPA & control mice. Inset in N shows an area with adipose tissue in Pdx1-Cre;HIF2dPA pancreata. Immunofluorescence of amylase & KRT19 shows no differences between Pdx1-Cre;HIF2dPA & control mice at P0 (G,H). Duct-like tubular structures & loss of amylase immunoreactivity in two- (K,L) & eight-week-old (O,P) Pdx1-Cre;HIF2dPA mice compared to control mice. Note areas with normal acini in 8-week-old Pdx1-Cre;HIF2dPA mice (white asterisk in O). Insets in (H,L & P) show higher magnification pictures. DAPI staining is shown in blue in (C,D,G,H,K,L,O & P). Scale bars = 50 µm for (C,D); 100 µm for (E–P). ***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.](http://images.novusbio.com/fullsize/nb100-122_rabbit-polyclonal-hif-2-alpha-epas1-antibody-310202415541931.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-479_rabbit-polyclonal-hif-1-alpha-antibody-310202415532013.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-479_rabbit-polyclonal-hif-1-alpha-antibody-310202415532077.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-479_rabbit-polyclonal-hif-1-alpha-antibody-31020241682177.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-479_rabbit-polyclonal-hif-1-alpha-antibody-310202415535154.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-479_rabbit-polyclonal-hif-1-alpha-antibody-310202415541975.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-479_rabbit-polyclonal-hif-1-alpha-antibody-310202415525255.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-479_rabbit-polyclonal-hif-1-alpha-antibody-310202415532080.jpg)
![Western Blot: HIF-1 alpha Antibody [NB100-449] - Morphological alterations induced by DMOG are HIF-1alpha dependent. glEND.2 cells were stably transfected with shGFP, shHIF-1alpha or shHIF-2alpha. The clones were cultured under normoxic (N) or hypoxic (H, 1% O2) conditions, or treated with DMOG for 6 h. 30 ug protein of cellular protein were loaded for the detection of HIF-1alpha, and nuclear extracts (20 ug protein) were used for the detection of HIF-2alpha. Image collected and cropped by CiteAb from the following publication (//biosignaling.biomedcentral.com/articles/10.1186/1478-811X-11-80) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Knockdown-Validated-NB100-449-img0050.jpg "<b>Genetic Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody [NB100-449] - Morphological alterations induced by DMOG are HIF-1alpha dependent. glEND.2 cells were stably transfected with shGFP, shHIF-1alpha or shHIF-2alpha. The clones were cultured under normoxic (N) or hypoxic (H, 1% O2) conditions, or treated with DMOG for 6 h. 30 ug protein of cellular protein were loaded for the detection of HIF-1alpha, and nuclear extracts (20 ug protein) were used for the detection of HIF-2alpha. Image collected and cropped by CiteAb from the following publication (//biosignaling.biomedcentral.com/articles/10.1186/1478-811X-11-80) licensed under a CC-BY license.")
![<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Western Blot: HIF-1 alpha Antibody [NB100-449] - HIF-1 alpha induction on Caki-1 cell lysate using CoCl2. Image from verified customer review.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Western-Blot-NB100-449-img0015.jpg)
![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.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Simple-Western-NB100-449-img0032.jpg)
![Immunohistochemistry: HIF-1 alpha Antibody [NB100-449] - Mouse Brain, Neurons 40X](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Immunohistochemistry-NB100-449-img0034.jpg)
![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](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Immunohistochemistry-NB100-449-img0047.jpg)
![<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).](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Western-Blot-NB100-449-img0038.jpg)
![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.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Western-Blot-NB100-449-img0053.jpg)
![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.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Immunocytochemistry-Immunofluorescence-NB100-449-img0039.jpg)
![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.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Immunohistochemistry-Paraffin-NB100-449-img0052.jpg)
![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.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Flow-Cytometry-NB100-449-img0035.jpg)
![<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.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Western-Blot-NB100-449-img0033.jpg)
![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).](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Western-Blot-NB100-449-img0036.jpg)
![Western Blot: HIF-1 alpha Antibody [NB100-449] - Detection of mouse HIF-1 alpha on hypoxia treated MEFs](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Western-Blot-NB100-449-img0037.jpg)
![<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.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Western-Blot-NB100-449-img0042.jpg)
![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.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Western-Blot-NB100-449-img0043.jpg)
![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).](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Western-Blot-NB100-449-img0044.jpg)
![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).](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Western-Blot-NB100-449-img0045.jpg)
![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.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Western-Blot-NB100-449-img0046.jpg)
![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.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Western-Blot-NB100-449-img0051.jpg)
![<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody [NB100-449] - Formaldehyde-fixed asynchronous HeLa cells.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Immunocytochemistry-Immunofluorescence-NB100-449-img0006.jpg)
![Immunoprecipitation: HIF-1 alpha Antibody [NB100-449] - HIF-1 alpha analysis in HEK293 cells. Image from verified customer review.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Immunoprecipitation-NB100-449-img0005.jpg)
![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.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Immunoprecipitation-NB100-449-img0048.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-310202415284545.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-310202415304212.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-310202415363719.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-31020241534344.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-310202415384135.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-310202415293325.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-310202415394514.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-310202415291950.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-310202415304256.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-31020241553383.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-310202415532021.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-310202415525284.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-31020241612685.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-31020241552391.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-31020241554543.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-31020241554544.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-31020241612613.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-31020241612635.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-3102024155453.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-31020241554521.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-31020241552522.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-31020241682187.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-31020241683944.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-31020241612654.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-310202415533837.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-31020241612628.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-310202415533827.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-31020241682146.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-31020241682155.jpg)
![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.](http://images.novusbio.com/fullsize/nb100-449_rabbit-polyclonal-hif-1-alpha-antibody-310202415541992.jpg)
![Simple Western: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Lane view shows a specific band for HIF-2 alpha in 0.5 mg/mL of Hypoxic HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/HIF-2-alpha-EPAS1-Antibody-ep190b-Simple-Western-NB100-132-img0009.jpg "<b>Biological Strategies Validation. </b>Simple Western: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Lane view shows a specific band for HIF-2 alpha in 0.5 mg/mL of Hypoxic HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.")
![<b>Biological Strategies Validation. </b>Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Analysis of HIF-2 alpha stabilization over time in 791T cells following exposure to hypoxia. Image using the HRP form of this antibody (NB100-132H). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23785417/) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/HIF-2-alpha-EPAS1-Antibody-ep190b-Western-Blot-NB100-132-img0017.jpg)
![Immunohistochemistry-Paraffin: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Analysis of HIF-2 in human cardiac myocytes using HIF-2 alpha/EPAS1 Antibody (ep190b).](http://images.novusbio.com/fullsize/HIF-2-alpha-EPAS1-Antibody-ep190b-Immunohistochemistry-Paraffin-NB100-132-img0018.jpg)
![Flow Cytometry: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - HIF-2 alpha antibody was tested at 1:400 in HepG2 cells using an Alexa Fluor 488 secondary (shown in purple). M1 is defined by unstained cells.](http://images.novusbio.com/fullsize/HIF-2-alpha-EPAS1-Antibody-ep190b-Flow-Cytometry-NB100-132-img0006.jpg)
![Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Mouse aortic endothelial cells treated (1%) or not treated (20.9%) in hypoxia for 3 hrs. Cells where also transfected with a specific siRNA against (siHIF-2) or a control siRNA (-). Western blot image submitted by a verified customer review.](http://images.novusbio.com/fullsize/HIF-2-alpha-EPAS1-Antibody-ep190b-Western-Blot-NB100-132-img0024.jpg)
![Immunohistochemistry-Paraffin: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Renal tubule specific models of Vhl deletion. Histological images of representative renal sections from 12 month old control, Pax8-CreERT2/Vhldelta/delta and Slc22a6-CreERT2/Vhldelta/delta mice (stains and antibodies as indicated, arrowheads indicate abnormal vascularization). Scale bars, 100 um. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0148055), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/HIF-2-alpha-EPAS1-Antibody-ep190b-Immunohistochemistry-Paraffin-NB100-132-img0022.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Analysis of HepG2 without Cobalt (II) Chloride (1), HepG2 with Cobalt (II) Chloride (2), HepG2 normoxic (3), HepG2 hypoxic (4), HepG2 without Cobalt (II) Chloride (5), HepG2 with Cobalt (II) Chloride (6), HepG2 normoxic (7), and HepG2 hypoxic (8) using this antibody (NB100-132) at 1 - 2 ug/mL.](http://images.novusbio.com/fullsize/HIF-2-alpha-EPAS1-Antibody-ep190b-Western-Blot-NB100-132-img0007.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - [HRP] [NB100-132H] - Analysis of HIF-2 alpha stabilization over time in HOS cells following exposure to hypoxia. Image using the HRP form of this antibody (NB100-132H). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23785417/) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/HIF-2-alpha-EPAS1-Antibody-ep190b-Western-Blot-NB100-132-img0019.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Western blot analysis and quantification of HIF-2 alpha expression in the cortex 4.5 hours after lipopolysaccharide (LPS) administration for all LPS groups and the control group (SHAM, white bar). With the exception of the vagotomy group (VGX LPS, gray bar), no significant differences to the SHAM group were found in the LPS-treated and sham-operated (SHAM+LPS, light gray bar) or vagus nerve-stimulated groups (VGX LPS+STIM, black bar). The significant increase in the VGX LPS (gray bar) group is an indicator of a hypoxic condition; * P<0.05 compared to SHAM; n=6 rats each. Data are given as the mean+/-SEM. Image collected and cropped by CiteAb from the following publication (//jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094-9-183), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/HIF-2-alpha-EPAS1-Antibody-ep190b-Western-Blot-NB100-132-img0020.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - 4-OOHCPA exposure induced HIF-2 alpha/EPAS1. A: Western blot analysis of HIF-2 alpha/EPAS1 protein (118KD) and actin (43KD) in limbs at 10 min, 1, 3, 6, and 24 h after treatment with 4-OOHCPA at 0.3 ug/mL (L) 1.0 ug/mL (M) or 3.0 ug/mL (H). P represents the positive control. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0051937), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/HIF-2-alpha-EPAS1-Antibody-ep190b-Western-Blot-NB100-132-img0021.jpg)
![Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Analysis of HIF-2 alpha in human endometrium using. Donkey anti-mouse Alexa Fluor 488 secondary antibody was used. IHC image submitted by a verified customer review.](http://images.novusbio.com/fullsize/HIF-2-alpha-EPAS1-Antibody-ep190b-Immunohistochemistry-NB100-132-img0016.jpg)
![<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Functional role of HIF2A in the transcriptional regulation of amphiregulin (AREG) in human cardiac myocytes. Immunoblot for HIF1A or HIF2A from shRNA-transfected normoxic or hypoxic HCM. Beta-Actin (ACTb) served as a loading control. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29483579/) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/HIF-2-alpha-EPAS1-Antibody-ep190b-Knockdown-Validated-NB100-132-img0025.jpg)
![Immunohistochemistry-Paraffin: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - HIF-1 alpha & HIF-2 alpha IHC signals in hypoxic areas of transgenic mouse mammary tumours.A, C Hypoxyprobe (Hyp.pr.) allows visualisation of hypoxic tumour areas. B. Hypoxic peri-necrotic HIF-1 alpha -positive cells display nuclear staining. D. HIF-2 alpha -positive cells show cytoplasmic staining with or without appreciable nuclear staining. * necrosis. E, F. IHC staining for HIF-1 alpha on MCF-7 breast cancer cells grown under control (E) & hypoxic (F) conditions respectively. G, H. Control (G) & hypoxic (H) MCF-7 cells IHC stained for HIF-2 alpha . 20x obj. Insets show magnification of the boxed area. Size bars 50 μm. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0125771), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-310202415371976.jpg)
![Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - HIF-2 alpha expression in the involuting mammary gland.Inserts are enlargements of the indicated areas. Size bars: 50 μm, 40x obj was used in all micrographs. A. In the early involuting gland, the morphology resembles the lactating gland & the basement membrane is evident at this stage. B-D. As tissue remodelling proceeds during involution, the collagen layer becomes unstructured. E-H. HIF-2 alpha -positive cells were detected at all studied stages of involution. I-L. Macrophage infiltration (F4/80 positive) was first evident at the fifth day of involution (J) & increased with time (K, L). Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0125771), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-310202415345323.jpg)
![Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Oxygen concentration-dependence of dioxygenase inhibition.A. HIF1 alpha , HIF2 alpha , & H3K9me2 abundance by western blotting in shRNA1 & shRNA2 cells incubated in 21%, 10%, or 2% oxygen for 72 h. Actin & total H3 serve as loading controls. B. HIF1 alpha , HIF2 alpha , & H3K9me2 abundance by western blotting in SDHC knockout iMEFs incubated in 21%, 10%, or 2% oxygen for 72 h. iMEFs were treated with 1 μM TAM for 7 d prior to analysis. (C-D) Rescue of succinate inhibition of JMHD & PHD inhibition using 0.25 mM octyl-alpha -ketoglutarate (octyl-alpha -KG) in SDHB knockdown HEK293 cells & SDHC knockout mouse iMEFs. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0127471), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-31020241533499.jpg)
![Immunohistochemistry-Paraffin: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Lactating mammary gland.Smaller panels display enlargements of the indicated areas. Size bars: 50 μm, 40x obj was used for all micrographs. A. Collagen I IHC allows visualisation of the basement membrane surrounding the dilated ducts. B. HIF-1 alpha was not detected in the epithelial cells of the lactating gland (compare with Fig 1). C. Macrophage infiltration was sparse in the lactating mammary gland as judged by F4/80 IHC. D. A subset of cuboidal luminal epithelial cells was distinctively positive for HIF-2 alpha . E. The percentage of HIF-2 alpha -positive out of total luminal epithelial cells was counted in sections from three mice. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0125771), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-310202415384198.jpg)
![Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Mainly CK14-positive, but not CK8-positive, mammary epithelial cells are HIF-2 alpha positive at day 14 post weaning.Left panels 20x & right 40x objective. Size bars 50 μm. a, A, b & B. Double IHC for HIF-2 alpha (green) & CK8 (a, A) & CK14 (b, B), respectively (red), reveal that few (if any) CK8-positive luminal cells are HIF-2 alpha positive. Numerous CK14-expressing cells, which include basal & stem/progenitor cells, were positive for HIF-2 alpha . c, C. F4/80 IHC was performed on an adjacent tissue section detect macrophages. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0125771), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-31020241535676.jpg)
![Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Renal tubule specific models of Vhl deletion.(A) PCR analysis of recombination at the Vhl locus in the kidneys of mice with combinations of Pax8-CreERT2, Slc22a6-CreERT2 & the Vhl floxed (fl) & wild-type (+) alleles. The positions of the bands representing the Vhl floxed, wild type (Wt) & recombined ( delta ) alleles are indicated. (B) Histological images of representative renal sections from 12 month old control, Pax8-CreERT2/Vhl delta / delta & Slc22a6-CreERT2/Vhl delta / delta mice (stains & antibodies as indicated, arrowheads indicate abnormal vascularisation). Scale bars, 100μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26866916), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-31020241539741.jpg)
![Immunohistochemistry-Paraffin: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - HIF1 alpha & H3K9me2 accumulation & 5-hydroxy-methyl-2’-deoxycytidine (5hmdC) depletion in PGL specimens compared to controls.Normal ganglia 1 (NG1), normal ganglia 2 (NG2) & IDH-mutant (IDH). Sporadic PGL (Spo. PGL). A. HIF1 alpha staining. B. HIF2 alpha staining. C. H3K9me2 staining. Arrows indicate H3K9me2 staining in nuclei of neurons or chief cells. D. 5hmdC staining. Arrows indicate 5hmdC staining in the nuclei of neurons & chief cells. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0127471), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-310202415345316.jpg)
![Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Localization of HIF2A immunoreactivity in limbs.HIF-2A reactivity was detected in control limbs at 1 & 3 h in the apical ectodermal ridge, interdigital area (I.R.) & developing cartilaginous anlagen (Digits). 4-OOHCPA exposure increased HIF2A immunoreactivity in the apical ectodermal ridge & interdigital regions (I.R.) at 3 h. No differences were observed between control & drug-treated limbs after 6 h or 24 h of culture. 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.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-310202415304236.jpg)
![Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Hypoxic repression of ER-alpha is dependent on HIF-1 alpha . a Representative western blots of HIF-1 alpha , HIF-2 alpha & beta -actin protein in ten ER-positive cell lines grown at normoxia or hypoxia (1% O2, 24 h). b qPCR analysis of HIF1A mRNA levels in MCF7, BT474, T47D & ZR75B transfected with shScramble or shHIF1A. Relative HIF1A mRNA levels normalized to TBP. (Change in HIF1A levels: *p < 0.0001 MCF7, *p = 0.012 BT474, *p < 0.0001 T47D, *p = 0.0046 ZR75B). c Representative western blots of HIF-1 alpha , ER-alpha & beta -actin protein from MCF7, BT474, T47D & ZR75B with either shScramble or shHIF1A at normoxia & hypoxia (1% O2, 24 h). beta -actin is used as a loading control. d Representative western blot of HIF-1 alpha , ER-alpha & beta -actin protein from MCF7, BT474, T47D & ZR75B with or without the transfection of stabilized HIF-1 alpha (HIF-1 alpha ODD) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28320353), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-3102024165744.jpg)
![Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - HIF-1 alpha & HIF-2 alpha expression in the virgin mammary gland.A. Virgin mammary glands (30 & 70 days old) showed no conspicuous basal membrane, as visualised by collagen IV IHC (a, b). There was also no detectable expression of HIF-2 alpha in the epithelial cells (c, d). Macrophages (F4/80 positive) were few. In panel c, a single HIF-2 alpha -positive cell was detected, & the adjacent F4/80 IHC section (e) suggested that this cell is a macrophage. B. Expression of HIF-1 alpha in mammary epithelium in the 70-day-old virgin mouse. Top panel, orientation slide with haematoxylin (HTX) staining, 20x obj. *lymph node. Panels b, d, f. Cross-section of a developing duct close to the invading tip at a stage where the lumen is not yet evacuated, 40x obj. Panels c, e, g. Cross-section of a less mature part of a duct, 40x obj. CK14-expressing cells (marker of basal mammary epithelial cells) can be seen in more than one cell layer (panels b & c, arrow-head). At this stage, the lumen is evacuated, but there is still more than one layer of epithelial cells. HIF-1 alpha IHC on the adjacent sections (panels d, e) showing nuclear expression in non-basal epithelial cells (highlighted by red arrows). Basal (CK14 positive) epithelial cells did not express HIF-1 alpha (black arrows). Mammary epithelial expression of HIF-2 alpha was not detected at these developmental stages. Size bars: 50 μm. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0125771), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-310202415532017.jpg)
![Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - HIF2 alpha inhibitor 76 suppressed CoCl2-induced immature phenotypic characteristics of BCSCs(A) The inhibitory effect of small molecule HIF2 alpha inhibitor 76 for 24h (10 μM for 4T1 cells; 25 μM for Hs578T cells) on CoCl2-induced expression of HIF2 alpha was assessed in both 4T1 & Hs578T cells by western blot analysis. (B) HIF2 alpha inhibitor 76 inhibited primary (with HIF2 alpha inhibitor 76 daily treatment) & second sphere formation (without additional HIF2 alpha inhibitor 76 treatment) in both 4T1 & Hs578T cells. The sizes of spheres greater than 100 μm were enumerated, with a representative image of a tumor-sphere shown. The data represents an average of three independent experiments. (C) Treatment of 4T1 & Hs578T cells with HIF2 alpha inhibitor 76 for 24 h led to a decrease in the percentage of CD44+/CD24−-positive cells as a proportion of total cancer cells. (D) 4T1 & Hs578T cells treated with CoCl2 (100 μM) for 24h & HIF2 alpha inhibitor 76 for 24h (10 μM for 4T1 cells; 25 μM for Hs578T cells) either alone or together were evaluated for the expression levels of stem cell markers c-Myc, Klf4, Oct4, & Nanog by Real-time PCR. Abbreviations: TSFE, Tumor sphere-forming efficiency. beta -actin was used as the internal control. The results represent the mean ± SD from three independent experiments. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.9846), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-310202415525281.jpg)
![Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Phd2 deletion leads to activation of the Akt–mTOR pathway. a RPPA analysis of tumors with homozygous deletion of Phd2. Tumor tissues from Tyr::CreER; BRafV600E; Phd2−/− or Tyr::CreER; BRafV600E mice were processed & analyzed by RPPA assays. The analyses identified proteins that were significantly changed in mouse melanomas compared to nevi. b Activation of Akt–mTOR pathway after phd2 deletion. Tumor tissues were processed & western blots showed stabilization of HIF-1 alpha & HIF-2 alpha proteins after Phd2 depletion. Increased phosphorylation of Akt, 4EBP1 & S6K was observed in tumors from Tyr::CreER; BRafV600E; Phd2−/− compared with those of Tyr::CreER; BRafV600E mice. c Re-expression of Phd2 inhibits the Akt–mTOR pathway. A BRafV600E; Phd2−/− mouse melanoma cell line was established from melanomas in Tyr::CreER; BRafCA; Phd2lox/lox mice. Phd2 was ectopically reintroduced in these tumor cells. Western blot analysis showed that degradation of HIF-1 alpha & HIF-2 alpha proteins with decreased expression of VEGFR2 decreased phosphorylation of Akt, 4EBP1 & S6K. d Pharmacological inhibition (FM19G11) of HIF pathway in BRafV600E; Phd2−/− melanoma cells. A similar but more pronounced inhibition of the Akt–mTOR pathway was observed using the HIF inhibitor. beta -Actin was used as a loading control. Results are representative of three independent experiments Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30575721), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-3102024161269.jpg)
![Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - BAY 87-2243 inhibits hypoxia-inducible factor (HIF-1 alpha ) & HIF-2 alpha protein accumulation in hypoxic H460 cells H460 under hypoxia but has no effect on HIF-1 alpha protein levels induced by hypoxia mimetics & has no effect on prolyl hydroxylase 2 (PHD2) activity. (A, B) H460 cells were cultured for 16 h under normoxia or hypoxia (1% pO2) in the absence or presence of various concentrations of BAY 87-2243. HIF-1 alpha (A) & HIF-2 alpha (B) protein levels were assessed by Western Blot in whole cell extracts. beta -actin was used as a loading control. (C) H460 cells were cultured for 16 h under normoxia with the PHDs desferrioxamine (DFO) & CoCl2 plus/minus BAY 87-2243 before the HIF1 alpha protein levels in cellular extracts were quantified by Western Blot. beta -actin was used to as a loading control. (D) Effect of BAY 87-2243 on the recombinant PHD2-mediated hydroxylation of HIF-1 alpha peptide over time was measured in a biochemical assay. Hydoxylated peptide was quantified after incubation with purified VBC complex labeled with europium using fluorescence as a readout. The known PHD inhibitor N-oxalylglycine served as a positive control. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24403227), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-31020241555221.jpg)
![Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Cytosol-restricted Acss2 is enzymatically active.(A) Acetylation of ectopic HA-tagged HIF-2 alpha detected by immunoblotting (IB) with anti-HA or anti- acetylated lysine antibodies following immunoprecipitation (IP) with anti-HA antibody in stably transformed HT1080 cells with knockdown of endogenous Acss2 & rescue with ectopic wild-type (WT) or cytosol-restricted mutant (CYT) Acss2 without or with an SV40 nuclear localization signal fused to the amino terminus. Studies were performed under hypoxia, low glucose, or acetate exposure for the indicated periods. (B) Acetate-dependent lipid synthesis measured by 14C-acetate incorporation in HT1080 stably-transformed cells producing control or Acss2 shRNA downstream of a luciferase cDNA cassette & expressing ectopic control, WT, CYT, SV40-WT, or SV40-CYT Acss2. Cells were incubated under (A) control, (B) hypoxic, or (C) low glucose conditions for 48 hr with labeling performed during the last 24 hr. Comparison of samples within a given condition was made by one-way ANOVA followed by Dunnett’s multiple comparisons test using control shRNA knockdown/control rescue as reference with decreased samples noted (*, P<0.05). All values are means with SD. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0190241), licensed under a CC0-1.0 license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-310202415525235.jpg)
![Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - In vitro effects of diclofenac on proliferation & MYC expression in the human melanoma cell line MelIm.The human melanoma cell line MelIm was incubated with different concentrations of diclofenac (A), aspirin (ASA, B), & NS-398 (C), respectively, & proliferation was determined after 24 h. Results represent the mean +/− standard deviation of 12 (diclofenac) & 3 (ASA, NS-398) independent experiments, respectively. (D) MelIm were incubated for 24 h with or without diclofenac. Apoptotic cells were stained with Annexin-V-FITC/ 7-AAD & analyzed by flow cytometry. Results represent the mean +/− standard deviation of 3 independent experiments. (E-G) MYC, STAT3, HIF1a & HIF2a protein expression were determined in cell lysates of MelIm incubated for 2 or 24 h with or without diclofenac (E,F) or ASA (G). The effect of diclofenac on MYC promoter activity was determined by transient transfection of a 2632-bp MYC promoter fragment (H). MelIm were transfected in 6-well-plates & diclofenac was added after 5 h. Luciferase activity was determined 24 h after transfection. Results represent the mean +/− standard deviation of 3 independent experiments. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0066987), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-310202415535152.jpg)
![Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Phd2 deletion leads to activation of the Akt–mTOR pathway. a RPPA analysis of tumors with homozygous deletion of Phd2. Tumor tissues from Tyr::CreER; BRafV600E; Phd2−/− or Tyr::CreER; BRafV600E mice were processed & analyzed by RPPA assays. The analyses identified proteins that were significantly changed in mouse melanomas compared to nevi. b Activation of Akt–mTOR pathway after phd2 deletion. Tumor tissues were processed & western blots showed stabilization of HIF-1 alpha & HIF-2 alpha proteins after Phd2 depletion. Increased phosphorylation of Akt, 4EBP1 & S6K was observed in tumors from Tyr::CreER; BRafV600E; Phd2−/− compared with those of Tyr::CreER; BRafV600E mice. c Re-expression of Phd2 inhibits the Akt–mTOR pathway. A BRafV600E; Phd2−/− mouse melanoma cell line was established from melanomas in Tyr::CreER; BRafCA; Phd2lox/lox mice. Phd2 was ectopically reintroduced in these tumor cells. Western blot analysis showed that degradation of HIF-1 alpha & HIF-2 alpha proteins with decreased expression of VEGFR2 decreased phosphorylation of Akt, 4EBP1 & S6K. d Pharmacological inhibition (FM19G11) of HIF pathway in BRafV600E; Phd2−/− melanoma cells. A similar but more pronounced inhibition of the Akt–mTOR pathway was observed using the HIF inhibitor. beta -Actin was used as a loading control. Results are representative of three independent experiments Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30575721), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-31020241555248.jpg)
![Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Phd2 deletion leads to activation of the Akt–mTOR pathway. a RPPA analysis of tumors with homozygous deletion of Phd2. Tumor tissues from Tyr::CreER; BRafV600E; Phd2−/− or Tyr::CreER; BRafV600E mice were processed & analyzed by RPPA assays. The analyses identified proteins that were significantly changed in mouse melanomas compared to nevi. b Activation of Akt–mTOR pathway after phd2 deletion. Tumor tissues were processed & western blots showed stabilization of HIF-1 alpha & HIF-2 alpha proteins after Phd2 depletion. Increased phosphorylation of Akt, 4EBP1 & S6K was observed in tumors from Tyr::CreER; BRafV600E; Phd2−/− compared with those of Tyr::CreER; BRafV600E mice. c Re-expression of Phd2 inhibits the Akt–mTOR pathway. A BRafV600E; Phd2−/− mouse melanoma cell line was established from melanomas in Tyr::CreER; BRafCA; Phd2lox/lox mice. Phd2 was ectopically reintroduced in these tumor cells. Western blot analysis showed that degradation of HIF-1 alpha & HIF-2 alpha proteins with decreased expression of VEGFR2 decreased phosphorylation of Akt, 4EBP1 & S6K. d Pharmacological inhibition (FM19G11) of HIF pathway in BRafV600E; Phd2−/− melanoma cells. A similar but more pronounced inhibition of the Akt–mTOR pathway was observed using the HIF inhibitor. beta -Actin was used as a loading control. Results are representative of three independent experiments Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30575721), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-310202415541965.jpg)
![Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - In-vivo lentiviral delivery of Cre-recombinase to renal tubular epithelium results in recombination of target genes.(a) Diagram of pCCIE lentiviral construct. Cre, Cre-recombinase; IRES, internal ribosome entry site; GFP, Green fluorescent protein. (b) Anti-VHL, HIF1a & GAPDH immunoblots of renal cortical protein lysates from Vhlwt/wt & Vhlfl/fl mice intrarenally injected with CCIE. Samples were collected 12 months post infection with each column representing an individual mouse. Blots were cropped to improve clarity, full-length blots are presented in Supplementary Fig. S3a. (c) Histological images of renal sections from Vhlwt/wt & Vhlfl/fl mice intrarenally injected with CCIE at 12 months post injection (stains & antibodies as indicated). Scale bars, 100 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26046460), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-310202415541994.jpg)
![Immunohistochemistry-Paraffin: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - HIF-1 alpha & HIF-2 alpha expression in the virgin mammary gland.A. Virgin mammary glands (30 & 70 days old) showed no conspicuous basal membrane, as visualised by collagen IV IHC (a, b). There was also no detectable expression of HIF-2 alpha in the epithelial cells (c, d). Macrophages (F4/80 positive) were few. In panel c, a single HIF-2 alpha -positive cell was detected, & the adjacent F4/80 IHC section (e) suggested that this cell is a macrophage. B. Expression of HIF-1 alpha in mammary epithelium in the 70-day-old virgin mouse. Top panel, orientation slide with haematoxylin (HTX) staining, 20x obj. *lymph node. Panels b, d, f. Cross-section of a developing duct close to the invading tip at a stage where the lumen is not yet evacuated, 40x obj. Panels c, e, g. Cross-section of a less mature part of a duct, 40x obj. CK14-expressing cells (marker of basal mammary epithelial cells) can be seen in more than one cell layer (panels b & c, arrow-head). At this stage, the lumen is evacuated, but there is still more than one layer of epithelial cells. HIF-1 alpha IHC on the adjacent sections (panels d, e) showing nuclear expression in non-basal epithelial cells (highlighted by red arrows). Basal (CK14 positive) epithelial cells did not express HIF-1 alpha (black arrows). Mammary epithelial expression of HIF-2 alpha was not detected at these developmental stages. Size bars: 50 μm. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0125771), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-31020241553385.jpg)
![Hypoxia inducible factor (HIF)‐2 alpha affects proliferation and self‐renewal capacity. A‐D, Embryo sections from trunk axial level. Proliferating EdU+ cells are green and electroporated cells (morpholinos, A; pCI‐CTRL and pCI‐EPAS1, C) are red. DAPI counterstains nuclei. Only construct targeted neural crest cells (above and outside of dotted line) were quantified (n = 82 [5′‐mispair morpholino] and n = 303 [EPAS1 morpholino], B); n = 211 (pCI‐CTRL) and n = 139 (pCI‐EPAS1), D. Statistical significance calculated using one‐way analysis of variance (ANOVA). E, Relative mRNA expression of EPAS1 in crestosphere cells established from cranial or trunk axial level measured by qRT‐PCR. Expression is presented as mean of n = 4 (cranial) or n = 3 (trunk) biological replicates and error bars represent SEM. Statistical significance calculated using two‐sided student's t test. F,G, Primary sphere assay of crestospheres established from embryos previously electroporated in ovo with non‐ (CTRL) or EPAS1 (EPAS1.1, F, EPAS1.2, G, targeting gRNAs. One cell/well, n = 10 wells/group). Number of spheres were manually counted after 1 week. Statistical significance was calculated by one‐way ANOVA. H, Size of spheres formed in, G. Manual measurements using the ImageJ software to convert to factual unit (μm). Statistical significance was determined by one‐way ANOVA. Inserted photographs of representative spheres from each group Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32940375), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-132_mouse-monoclonal-hif-2-alpha-epas1-antibody-ep190b-immunocytochemistry-immunofluorescence-13220261920712.jpg)
![ELISA HIF-1 alpha [Biotin]](http://images.novusbio.com/fullsize2/dyc1935-2_human-mouse-total-hif-1-alpha-duoset-ic-elisa-5-plate-elisa-10920209831.jpg "ELISA HIF-1 alpha [Biotin]")
![Western Blot: FIH-1/HIF-1AN Antibody [NB100-428] - Analysis of astrocytes lysate from Rat brain using FIH-1/HIF-1AN antibody at 1:500 dilution with ECL detection. The antibody detected a major band at ~40 kDa position representing FIH-1/HIF-1AN protein.](http://images.novusbio.com/fullsize/FIH-1-HIF-1AN-Antibody-Western-Blot-NB100-428-img0007.jpg "Western Blot: FIH-1/HIF-1AN Antibody [NB100-428] - Analysis of astrocytes lysate from Rat brain using FIH-1/HIF-1AN antibody at 1:500 dilution with ECL detection. The antibody detected a major band at ~40 kDa position representing FIH-1/HIF-1AN protein.")
![Western Blot: FIH-1/HIF-1AN Antibody [NB100-428] - Representative picture of western blot in histopathologically unchanged tissue (N) and primary cancerous tissue (C) from patients with CRC. Immunodetection of bands was performed with Rp anti- PHD1 (Catalog # NB100-310), - PHD2 (Catalog # NB100-137), - PHD3 (Catalog # NB100-139) and -FIH-1/HIF-1AN Ab (Catalog # NB100-139), followed by incubation with goat anti-rabbit HRP-conjugated Ab. The membrane was stripped and incubated with Rp anti-GAPDH Ab, followed by incubation with goat anti-rabbit HRP-conjugated Ab. Image collected and cropped by CiteAb from the following publication (//bmccancer.biomedcentral.com/articles/10.1186/1471-2407-13-526), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/FIH-1-HIF-1AN-Antibody-Western-Blot-NB100-428-img0010.jpg)
![Immunohistochemistry: FIH-1/HIF-1AN Antibody [NB100-428] - Detection of FIH-1 in rat brain tissue.](http://images.novusbio.com/fullsize/FIH-1-HIF-1AN-Antibody-Immunohistochemistry-NB100-428-img0008.jpg)
![Western Blot: FIH-1/HIF-1AN Antibody [NB100-428] - Rat VSMC (vascular smoot muscle) cells treated (siFIH) or not (CTL) with a specific siRNA against FIH-1. Western blot image submitted by a verified customer review.](http://images.novusbio.com/fullsize/FIH-1-HIF-1AN-Antibody-Western-Blot-NB100-428-img0009.jpg)
![<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Western Blot: FIH-1/HIF-1AN Antibody [NB100-428] - (C) Cytoplasmic FIH-1 (lower band) and MMP14 expression levels in both aerobic and hypoxic conditions in the 3 cell lines. (E) siRNA targeting FIH-1 was used to assess whether FIH-1 had any effect on CAIX expression in MCF-7 cells cultured in 0.5% O2 conditions. cont, control. m, transfection reagent alone. scr, scrambled siRNA. pool, mixture of 4 siRNAs targeting FIH-1. 21, single siRNA targeting FIH-1. (F) FIH-1 and CAIX expression levels in aerobic MCF-7 cells treated with either siRNA against FIH-1 alone, or both 400 uM CoCl2 and siRNA together for 24, 48 and 72h. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28476026/) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/FIH-1-HIF-1AN Antibody-Knockdown Validated-NB100-428-img0011.jpg)
![Immunohistochemistry: FIH-1/HIF-1AN Antibody [NB100-428] - 3D target protein expression in multicellular tumor spheroids3D expression analysis was carried out in the MCF-7, MDA-MB-231 & HBL-100 cell lines using multicellular tumor spheroids. The spheroids were cultured in spinner flasks for 1 week before fixation. Hypoxyprobe was used to examine the formation of hypoxic areas within the spheroids. NHE1, CAIX, MMP14 & FIH-1 expression was also analyzed. Incubation for an hour with PBS instead of primary antibody acted as a control. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.17143), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-428_rabbit-polyclonal-fih-1-hif-1an-antibody-310202415384122.jpg)
![Western Blot: HIF-1 alpha Antibody Pack [NB100-905] - HIF-1 alpha induction by CoCl2 on Caki-1 cell lysate. Image from verified customer review. HIF-1 alpha Antibody (H1alpha67) [NB100-105]](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Pack-Western-Blot-NB100-905-img0048.jpg "<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody Pack [NB100-905] - HIF-1 alpha induction by CoCl2 on Caki-1 cell lysate. Image from verified customer review. HIF-1 alpha Antibody (H1alpha67) [NB100-105]")
![<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody Pack [NB100-905] - 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. HIF-1 alpha Antibody (H1alpha67) [NB100-105]](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Pack-Knockout-Validated-NB100-905-img0049.jpg)
![Chromatin Immunoprecipitation: HIF-1 alpha Antibody Pack [NB100-905] - 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. HIF-1 alpha Antibody [NB100-479]](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Pack-Chromatin-Immunoprecipitation-NB100-905-img0041.jpg)
![Simple Western: HIF-1 alpha Antibody Pack [NB100-905] - 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. HIF-1 alpha Antibody [NB100-449]](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Pack-Simple-Western-NB100-905-img0042.jpg)
![Immunohistochemistry: HIF-1 alpha Antibody Pack [NB100-905] - HIF-1alpha expression in PDAC and adjacent normal pancreatic tissue. PDAC with weak (intensity 1) nuclear and cytoplasmic HIF-1alpha staining (left). PDAC with strong (intensity 3) nuclear and moderate (intensity 2) cytoplasmic HIF-1alpha staining (right). Image collected and cropped by CiteAb from the following publication (//wjso.biomedcentral.com/articles/10.1186/s12957-018-1432-4), licensed under a CC-BY license. HIF-1 alpha Antibody (H1alpha67) [NB100-105]](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Pack-Immunohistochemistry-NB100-905-img0051.jpg)
![Flow Cytometry: HIF-1 alpha Antibody Pack [NB100-905] - Analysis using the Alexa Fluor (R) 488 conjugate of NB100-105. Staining of HIF-1 alpha in multiple myeloma cells: H929 cells (0.5x10^6) 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. HIF-1 alpha Antibody (H1alpha67) [NB100-105]](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Pack-Flow-Cytometry-NB100-905-img0050.jpg)
![<b>Biological Strategies Validation, Genetic Strategies Validation and Orthogonal Strategies Validation.</b>Western Blot: HIF-1 alpha Antibody Pack [NB100-905] - Effects of in vitro hypoxia on the expression of mouse HIF-1 alpha in HIF-1 alpha+/- and HIF-1 alpha+/+ astrocytes. Confluent astrocyte monolayers of both cell types were exposed to a 6 h in vitro hypoxia. HIF-1 alpha mRNA expression was determined by RT-PCR as described in Materials and Methods. Each bar represents the mean +/- SD of relative density/volumes of the bands on film negatives from at least three experiments. Asterisks and number sign indicate significant difference (p < 0.01; one-way ANOVA, followed by multiple comparisons among means). Image collected and cropped by CiteAb from the following publication (//jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094-4-12), licensed under a CC-BY license. HIF-1 alpha Antibody [NB100-654]](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Pack-Western-Blot-NB100-905-img0053.jpg)
![Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody Pack [NB100-905] - Detection of HIF-1 alpha (red dye 568) in a cultured raw mouse macrophage cell line, using NB100-131. 100X magnification. Photos courtesy of Susan Alexander and Hattie Gresham, PhD. HIF-1 alpha Antibody (ESEE122) [NB100-131]](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Pack-Immunocytochemistry-Immunofluorescence-NB100-905-img0037.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody Pack [NB100-905] - 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. Also shown are Western blots of nuclear HIF-1alpha at the time of irradiation. Corresponding Western blots of lamin A/C are shown as a loading control. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0111199) licensed under a CC-BY license. HIF-1 alpha Antibody [NB100-479].](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Pack-Western-Blot-NB100-905-img0040.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody Pack [NB100-905] - HIF-1 alpha induction on Caki-1 cell lysate using CoCl2. Image from verified customer review. HIF-1 alpha Antibody [NB100-449]](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Pack-Western-Blot-NB100-905-img0044.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody Pack [NB100-905] - Effects of CoCl2 treatment on HIF-1 alpha expression in mouse astrocytes. Western blots using nuclear proteins show that both hypoxia (H) and CoCl2 (Co) up-regulated HIF-1 alpha protein in HIF-1 alpha+/+ and HIF-1 alpha+/- cells. There was no HIF-1 alpha protein detected in control cells. Image collected and cropped by CiteAb from the following publication (//jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094-4-12), licensed under a CC-BY license. HIF-1 alpha Antibody [NB100-654]](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Pack-Western-Blot-NB100-905-img0052.jpg)
![Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody Pack [NB100-905] - Detection of HIF-1 alpha (red dye) in a cell cytospin from a lavage of a murine skin pouch infected with Staph Aureus, using NB100-131. 100X magnification. Blue dye is DAPI nuclea](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Pack-Immunocytochemistry-Immunofluorescence-NB100-905-img0035.jpg)
![Immunohistochemistry: HIF-1 alpha Antibody Pack [NB100-905] - Staining in normal mouse kidney showing (A) staining with antibody and (B) blocking peptide used at 10-fold excess concentration. Images provided by Dr Yves Heremans. Please see the product review of NBC1-18422 for additional details. HIF-1 alpha Antibody [NB100-479]](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Pack-Immunohistochemistry-NB100-905-img0038.jpg)
![Immunohistochemistry: HIF-1 alpha Antibody Pack [NB100-905] - Hif1alpha expression in preimplantation embryos. Co-staining of OCT4 and Hif1alpha in inner cell mass of blastocyst (top) and negative control images without primary antibodies (bottom). Scale bar: 30um. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0154309) licensed under a CC-BY license. HIF-1 alpha Antibody [NB100-479]](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Pack-Immunohistochemistry-NB100-905-img0039.jpg)
![Immunohistochemistry: HIF-1 alpha Antibody Pack [NB100-905] - Immunohistochemical analysis in non-GIST STS representing negative, and score 1-3 of Carbonic Anhydrase IX/CA9, GLUT-1, HIF-1 alpha, and HIF-2 alpha/EPAS1. non-GIST STS: non-gastrointestinal stromal tumor soft-tissue sarcomas, Carbonic Anhydrase IX/CA9: carbonic anhydrase IX, GLUT-1: glucose transporter-1, and HIF-1/2alpha: hypoxia induced factor 1/2alpha. Image collected and cropped by CiteAb from the following publication (//www.hindawi.com/journals/sarcoma/2012/541650/), licensed under a CC-BY license. HIF-1 alpha Antibody (ESEE122) [NB100-131]](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Pack-Immunohistochemistry-NB100-905-img0045.jpg)
![Immunohistochemistry: HIF-1 alpha Antibody Pack [NB100-905] - Analysis of HIF-1 alpha in paraffin-embedded mouse kidney tissue section using anti-HIF-1 alpha antibody. Image from verified customer review. HIF-1 alpha Antibody (ESEE122) [NB100-131]](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Pack-Immunohistochemistry-NB100-905-img0047.jpg)
![<b>Genetic Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody Pack [NB100-905] - Morphological alterations induced by DMOG are HIF-1alpha dependent. glEND.2 cells were stably transfected with shGFP, shHIF-1alpha or shHIF-2alpha. The clones were cultured under normoxic (N) or hypoxic (H, 1% O2) conditions, or treated with DMOG for 6 h. 30 ug protein of cellular protein were loaded for the detection of HIF-1alpha, and nuclear extracts (20 ug protein) were used for the detection of HIF-2alpha. Image collected and cropped by CiteAb from the following publication (//biosignaling.biomedcentral.com/articles/10.1186/1478-811X-11-80) licensed under a CC-BY license. HIF-1 alpha Antibody [NB100-449]](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-Pack-Knockdown-Validated-NB100-905-img0043.jpg)
![Western Blot: HIF-1 alpha Antibody (2443C) [NBP2-75978] - Total protein from Human U87 cells treated with or without CoCl2 and under Normoxic and Hypoxic conditions was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-HIF-1 alpha in 5% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-2443C-Western-Blot-NBP2-75978-img0004.jpg "<b>Biological Strategies Validation. </b>Western Blot: HIF-1 alpha Antibody (2443C) [NBP2-75978] - Total protein from Human U87 cells treated with or without CoCl2 and under Normoxic and Hypoxic conditions was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-HIF-1 alpha in 5% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.")
![<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Western Blot: HIF-1 alpha Antibody (2443C) [NBP2-75978] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and HIF-1 alpha knockout HeLa cell line (KO) untreated (-) or treated (+) with 1 mM DFO for overnight. PVDF membrane was probed with 2 ug/mL of Rabbit Anti-Human HIF-1 alpha Monoclonal Antibody (Catalog # NBP2-75978) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for HIF-1 alpha at approximately 120 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # MAB5718) is shown as a loading control.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-2443C-Knockout-Validated-NBP2-75978-img0006.jpg)
![Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody (2443C) [NBP2-75978] - Immunofluorescent staining of HIF-1 alpha using catalog number NBP2-75977 (clone 2443B). Staining was observed in Hela cells treated with DFO but not in untreated Hela cells.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-2443C-Immunocytochemistry-Immunofluorescence-NBP2-75978-img0002.jpg)
![Immunohistochemistry-Paraffin: HIF-1 alpha Antibody (2443C) [NBP2-75978] - HIF-1 alpha (clone 2443C) was detected in immersion fixed paraffin-embedded sections of human kidney using Rabbit Anti-Human HIF-1 alpha Monoclonal Antibody (Catalog # NBP2-75978) at 1.0 ug/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte (TM) HRP Polymer Antibody (Catalog # VC002). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the nuclei.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-2443C-Immunohistochemistry-Paraffin-NBP2-75978-img0003.jpg)
![Simple Western: HIF-1 alpha Antibody (2443C) [NBP2-75978] - Lane view shows a specific band for HIF-1 alpha in HepG2 +/- DFO cell lysate using 20 ug/mL. Electropherogram image of corresponding Simple Western lane view at WES molecular weight of 116 KDa. Image from an internal validation.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-2443C-Simple-Western-NBP2-75978-img0007.jpg)
![Simple Western: HIF-1 alpha Antibody (2443C) [NBP2-75978] - 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 NBP2-75978 (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.](http://images.novusbio.com/fullsize/HIF-1-alpha-Antibody-2443C-Simple-Western-NBP2-75978-img0008.jpg)
![SDS-Page: Recombinant Human HIF-1 alpha Protein [H00003091-P01] - 12.5% SDS-PAGE Stained with Coomassie Blue.](http://images.novusbio.com/fullsize/Recombinant-Human-HIF-1-alpha-Protein-SDS-Page-H00003091-P01-img0002.jpg "SDS-Page: Recombinant Human HIF-1 alpha Protein [H00003091-P01] - 12.5% SDS-PAGE Stained with Coomassie Blue.")
![Western Blot: HIF-1 alpha Antibody - Exon 13 [NBP1-47181] - Human recombinant HIF-1 alpha protein.](http://images.novusbio.com/fullsize/HIF-1-alpha-[exon-13]-Antibody-Western-Blot-NBP1-47181-img0002.jpg "Western Blot: HIF-1 alpha Antibody - Exon 13 [NBP1-47181] - Human recombinant HIF-1 alpha protein.")