HIF-2 alpha/EPAS1: Novus Biologicals

NB100-122

HIF-2 alpha/EPAS1 Antibody - BSA Free

(37 Reviews)

(876 Publications)

0.1 ml

NB100-122

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

HIF-2-alpha-EPAS1-Antibody-Immunohistochemistry-NB100-122-img0028.jpg

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

<b>Genetic Strategies Validation. </b>HIF-2-alpha-EPAS1-Antibody-Knockout-Validated-NB100-122-img0035.jpg

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

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

<b>Biological Strategies Validation. </b>HIF-2-alpha-EPAS1-Antibody-Western-Blot-NB100-122-img0030.jpg

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

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

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

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

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

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

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

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

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.

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.

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.

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.

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.

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.

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 < 0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30209343), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: HIF-2 alpha/EPAS1 Antibody - BSA Free [NB100-122] - QSCs are hypoxic in the niche & express HIF2A, but not HIF1A.(A) Timeline of in vivo pimonidazole labeling in SC-INTACT mice & representative confocal images of uninjured/resting EDL myofibers (n >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.

Reactivity:

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

Species Glossary

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

Applications:

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

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

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-122AF350
Alexa Fluor 405	NB100-122AF405
Alexa Fluor 488	NB100-122AF488
Alexa Fluor 532	NB100-122AF532
Alexa Fluor 594	NB100-122AF594
Alexa Fluor 647	NB100-122AF647
Alexa Fluor 700	NB100-122AF700
Alexa Fluor 750	NB100-122AF750
Alexa Fluor Plus 405	NB100-122AFP405
Alexa Fluor Plus 488	NB100-122AFP488
Alexa Fluor Plus 555	NB100-122AFP555
Alexa Fluor Plus 594	NB100-122AFP594
Alexa Fluor Plus 647	NB100-122AFP647
Alexa Fluor Plus 680	NB100-122AFP680
Allophycocyanin	NB100-122APC
Allophycocyanin/Cy7	NB100-122APCCY7
Biotin	NB100-122B
CoraFluor 1	NB100-122CL1
DyLight 350	NB100-122UV
DyLight 405	NB100-122V
DyLight 488	NB100-122G
DyLight 550	NB100-122R
DyLight 594	NB100-122DL594
DyLight 650	NB100-122C
DyLight 680	NB100-122FR
DyLight 755	NB100-122IR
FITC	NB100-122F
HRP	NB100-122H
Janelia Fluor 525	NB100-122JF525
Janelia Fluor 549	NB100-122JF549
Janelia Fluor 646	NB100-122JF646
Janelia Fluor 669	NB100-122JF669
PE	NB100-122PE
PE/Atto594	NB100-122PEATT594
PE/Cy5.5	NB100-122PECY55
PE/Cy7	NB100-122PECY7
PerCP	NB100-122PCP
mFluor Violet 450 SE	NB100-122MFV450
mFluor Violet 500 SE	NB100-122MFV500
mFluor Violet 610 SE	NB100-122MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-122SS

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NB100-132

HIF-2 alpha/EPAS1 Antibody (ep190b)

(9 Reviews)

(195 Publications)

0.1 ml

NB100-132

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).

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.

<b>Biological Strategies Validation. </b>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.

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.

<b>Biological Strategies Validation. </b>HIF-2-alpha-EPAS1-Antibody-ep190b-Western-Blot-NB100-132-img0019.jpg

<b>Biological Strategies Validation. </b>HIF-2-alpha-EPAS1-Antibody-ep190b-Western-Blot-NB100-132-img0020.jpg

<b>Biological Strategies Validation. </b>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.

<b>Biological Strategies Validation and Genetic Strategies Validation.</b>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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

Hypoxia inducible factor (HIF)‐2 alpha is expressed in trunk neural crest cells. A,C, Immunostaining of HIF‐2 alpha in sections from trunk axial level of wild‐type chick embryos at HH11, A, HH13, B, and HH18, C. Arrow denotes ventrally migrating HIF‐2 alpha positive cells. D, Co‐immunostaining of HIF‐2 alpha and HNK1 (marker of migrating neural crest) in sections from trunk axial level of wild‐type HH18 chick embryos. Arrows denote migrating cells double positive for the two proteins. E, Sections of HH13 wild‐type embryo immunostained with DAPI for visualization of nuclei and secondary antibody only (donkey anti‐rabbit Alexa Fluor‐546). F, Western blot analysis for detection of HIF‐2 alpha protein at 21% and 1% O2 following siRNA mediated knockdown of HIF‐1 alpha or HIF‐2 alpha . DIP treated cells were used as a positive control and SDHA as loading control. Lanes between 21% and 1% siCTRL were removed from this figure, indicated by the black line. G, Immunohistochemical staining for HIF‐2 alpha in sections of SK‐N‐BE(2)c neuroblastoma cells cultured at 21% or 1% O2. H, Schematic of where oxygen measurements were performed. I, Oxygen saturation in the trunk of chick embryos during development measured ex ovo using microsensor technique. Error bars represent SEM, n ≥ 3 biologically independent replicates for each time point 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.

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.

Reactivity:

Hu, Mu, Rt, Bv, Ha

Species Glossary

Reactivity: Hu, Mu, Rt, Bv, Ha

Applications:

, , , , , , , , , , ,

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

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-132AF350
Alexa Fluor 405	NB100-132AF405
Alexa Fluor 488	NB100-132AF488
Alexa Fluor 532	NB100-132AF532
Alexa Fluor 594	NB100-132AF594
Alexa Fluor 647	NB100-132AF647
Alexa Fluor 700	NB100-132AF700
Alexa Fluor 750	NB100-132AF750
Allophycocyanin	NB100-132APC
Biotin	NB100-132B
CoraFluor 1	NB100-132CL1
DyLight 350	NB100-132UV
DyLight 405	NB100-132V
DyLight 488	NB100-132G
DyLight 550	NB100-132R
DyLight 594	NB100-132DL594
DyLight 650	NB100-132C
DyLight 680	NB100-132FR
DyLight 755	NB100-132IR
FITC	NB100-132F
HRP	NB100-132H
Janelia Fluor 525	NB100-132JF525
Janelia Fluor 549	NB100-132JF549
Janelia Fluor 646	NB100-132JF646
Janelia Fluor 669	NB100-132JF669
PE	NB100-132PE
PerCP	NB100-132PCP
mFluor Violet 450 SE	NB100-132MFV450
mFluor Violet 500 SE	NB100-132MFV500
mFluor Violet 610 SE	NB100-132MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Azide and BSA Free	NBP2-80766

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AF2997

HIF-2 alpha/EPAS1 Antibody [Unconjugated]

(36 Publications)

100 μg

AF2997

CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei in cancer cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." class="big_lightbox">

HIF-2a/EPAS1 was detected in immersion fixed paraffin-embedded sections of human prostate cancer tissue using Goat Anti-Human/Mouse/Rat HIF-2a/EPAS1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2997) at 0.3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # <a class=

CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei in cancer cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." alt="HIF-2a/EPAS1 was detected in immersion fixed paraffin-embedded sections of human prostate cancer tissue using Goat Anti-Human/Mouse/Rat HIF-2a/EPAS1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2997) at 0.3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei in cancer cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." class="big_thumb" />

CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei in spermatocytes. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections." class="big_lightbox">

HIF-2a/EPAS1 was detected in perfusion fixed frozen sections of mouse testis using Goat Anti-Human/Mouse/Rat HIF-2a/EPAS1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2997) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # <a class=

CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei in spermatocytes. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections." class="big_thumb" />

HAF109). A specific band was detected for HIF-2a/EPAS1 at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox">

<b>Biological Strategies Validation. </b>Western blot shows lysates of MCF-7 human breast cancer cell line, bEnd.3 mouse endothelioma cell line, and PC-12 rat adrenal pheochromocytoma cell line untreated (-) or treated (+) with 150 µM CoCl<SUB>2</SUB>for 16 hours. PVDF Membrane was probed with 1 µg/mL of Goat Anti-Mouse HIF-2a/EPAS1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2997) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # <a class=

HAF109). A specific band was detected for HIF-2a/EPAS1 at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

Kidney-derived Sca-1+ cells produce Epo. (A) Epo protein levels in supernatants obtained from Sca-1+ cells cultured in normoxia or hypoxia (n = 3 biological replicates). (B) HIF-1 alpha and HIF-2 alpha protein levels in Sca-1+ kidney-derived cells after exposure to normoxia (20% O2) or hypoxia (1% O2). Numbers below the blots indicate the fold change of the ratio of HIF-1 alpha /Tubulin and HIF-2 alpha /Tubulin in 1% O2 to the respective 20% O2 control. (C) Epo RNA levels in Sca-1+ cells that were incubated either at 20% O2 of 1% O2 as indicated (n = 4 biological replicates). (D) Mesenchymal stem cell markers in renal Sca-1+ cells on day 21 of culture; reads per kilobase million (RPKM). (E) Volcano plot of 4592 significantly up- and 5376 significantly downregulated mRNAs in kidney-derived Sca-1+ cells on day 21 in culture compared to day 0. Mean values ± SEM are shown. * p < 0.05. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35203399), licensed under a CC-BY license. Not internally tested by R&D Systems.

Hypoxia upregulates AK4 in an HIF-1 alpha -dependent manner. (A) Representative Western blot for HIF-1 alpha and AK4 followed by densitometric quantification of relative expression in PASMCs treated with 50 µM DFO for 24 h, compared to untreated control; n = 4. (B) Representative Western blot for HIF-1 alpha and AK4 and densitometric quantification of relative expression in PASMCs transfected with HIF-1 alpha siRNA (siHIF-1 alpha ), followed by hypoxic (HOX, 1% O2) exposure for 24 h, compared to non-targeting siRNA as a control (scrambled siRNA = scramble) under NOX or HOX; n = 3. (C,E) Relative mRNA expression of AK4 in PASMCs transfected with siHIF-1 alpha (C) or siEPAS1 (E), followed by hypoxic (HOX, 1% O2) exposure for 24 h, compared to control (scramble); n = 3–5. (D) Representative Western blot for EPAS1 (HIF-2 alpha ) and AK4 and densitometric quantification of relative expression in PASMCs transfected with EPAS1 siRNA (siEPAS1), followed by hypoxic (HOX, 1% O2) exposure for 24 h, compared to control (scramble) under NOX or HOX; n = 4. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p< 0.0001, (A,C,E) unpaired Student’s t-test or (B,D) one-way ANOVA followed by Tukey multiple comparisons test. Image collected and cropped by CiteAb from the following open publication (//www.mdpi.com/1422-0067/22/19/10371), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

,

Applications: ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	AF2997U-100UG
Alexa Fluor 405	AF2997V-100UG
Alexa Fluor 488	AF2997G-100UG
Alexa Fluor 532	AF2997X-100UG
Alexa Fluor 594	AF2997T-100UG
Alexa Fluor 647	AF2997R-100UG
Alexa Fluor 700	AF2997N-100UG
Alexa Fluor 750	AF2997S-100UG
Alexa Fluor Plus 405	AF2997AFP405-100UG
Alexa Fluor Plus 488	AF2997AFP488-100UG
Alexa Fluor Plus 555	AF2997AFP555-100UG
Alexa Fluor Plus 594	AF2997AFP594-100UG
Alexa Fluor Plus 647	AF2997AFP647-100UG
Alexa Fluor Plus 680	AF2997AFP680-100UG
Allophycocyanin	AF2997A
Biotin	AF2997B
CoraFluor 1	AF2997CL1
DyLight 350	AF2997D
DyLight 405	AF2997E
DyLight 488	AF2997K
DyLight 550	AF2997L
DyLight 594	AF2997M
DyLight 650	AF2997W
DyLight 680	AF2997Y
DyLight 755	AF2997Z
FITC	AF2997F
HRP	AF2997H
Janelia Fluor 525	AF2997JF525
Janelia Fluor 549	AF2997I
Janelia Fluor 646	AF2997J
Janelia Fluor 669	AF2997JF669
PE	AF2997P
PerCP	AF2997C
mFluor Violet 450 SE	AF2997MFV450
mFluor Violet 500 SE	AF2997MFV500
mFluor Violet 610 SE	AF2997MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF2997-SP

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NB100-902HIF2

HIF-2 alpha/EPAS1 Antibody Pack

(1 Review)

(12 Publications)

4 Vials

NB100-902HIF2

Simple Western: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2] - 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. HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132]

<b>Genetic Strategies Validation. </b>Western Blot: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2] - HIF-1alpha is the predominant transcriptional regulator of WNT11 expression during hypoxia. EMSCs isolated from the indicated mouse genotypes were infected with lentivirus expressing GFP or Cre recombinase. Non-infected cells and GFP infected cells served as controls. Immunoblot analyses of EMSCs derived from the indicated genotypes treated with 0.1 mM DMOG for 24 hrs. Attenuated WNT11 expression in Hif-1alpha KO EMSCs (lenti-Cre infected Hif-1af/f). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep21520) licensed under a CC-BY license. HIF-2 alpha/EPAS1 Antibody [NB100-122]

<b>Orthogonal Strategies Validation. </b>Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2] - Formalin-fixed paraffin-embedded tissue sections of human placenta were probed for HIF-2 alpha/EPAS1 mRNA (ACD RNAScope Probe, catalog #410598; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using Rabbit Polyclonal (Novus Biologicals catalog # NB100-122) at 1:100 dilution with one-hour incubation at room temperature followed by incubation with anti-rabbit IgG VisUCyte HRP Polymer Antibody (Catalog # VC003) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to trophoblastic cells. HIF-2 alpha/EPAS1 Antibody [NB100-122]

Immunohistochemistry: HIF-2 alpha/EPAS1 Antibody Pack [NB100-902HIF2] - Normal ganglia 1 (NG1), normal ganglia 2 (NG2) and IDH-mutant (IDH). Sporadic PGL (Spo. PGL). HIF2 alpha staining. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0127471) licensed under a CC-BY license. HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132]

Reactivity:

Hu, Mu, Rt, Ha, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, Ha, Rb

Applications:

, , , , , , , ,

Applications: , , , , , , , ,

Host:

Mouse

Host: Mouse

Validated by:

Validated by:

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AF2886

HIF-2 alpha/EPAS1 Antibody

(11 Publications)

100 μg

AF2886

HAF109). A specific band was detected for HIF-2a/EPAS1 at approximately 115 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox">

<b>Biological Strategies Validation. </b>Western blot shows lysates of MCF-7 human breast cancer cell line and PC-12 rat adrenal pheochromocytoma cell line untreated (-) or treated (+) with 150 µM CoCl<sub>2</sub>for 16 h. PVDF membrane was probed with 1 µg/mL of Human/Rat HIF-2a/EPAS1 Polyclonal Antibody, followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # <a class=

HAF109). A specific band was detected for HIF-2a/EPAS1 at approximately 115 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of MCF-7 human breast cancer cell line and PC-12 rat adrenal pheochromocytoma cell line untreated (-) or treated (+) with 150 µM CoCl₂for 16 h. PVDF membrane was probed with 1 µg/mL of Human/Rat HIF-2a/EPAS1 Polyclonal Antibody, followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for HIF-2a/EPAS1 at approximately 115 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. Theoct-3/4promoter was detected by standard PCR." class="big_lightbox">

<b>Biological Strategies Validation. </b>BG01V human embryonic stem cells treated with 150 µM CoCl2 overnight were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. HIF-2a/EPAS1/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human/Rat HIF-2a/EPAS1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2886) or control antibody (Catalog # <a class=

AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. Theoct-3/4promoter was detected by standard PCR." class="big_thumb" />

The DU145FP cells express higher levels of cancer stemness related genes. (A) Transcriptome analyses of prostate cancer stemness-related markers. The Log2 transformed ratio of FPKM values (e.g., FP/WT) are indicated by color-coded index bars. (B) Cells were stained with fluorophore conjugated monoclonal antibodies against CD44 and EPCAM, the antibody against integrin alpha 2 beta 3 without fluorophore conjugation was detected by using secondary antibody conjugated with fluorophore, and stained cells were analyzed by flow cytometry. The scatter plots and histograms show representative results. CTRL denote cell stained with isotype control antibody or only secondary antibody; ST denote cell stained with antibody against EPCAM and integrin alpha 2 beta 3; US denote unstained cells. (C) Immunoblotting confirmation of the protein expression of A-tubulin, GAPDH, SOX9, SLUG, HIF1A, HIF2A, CD44 and integrin alpha 2 beta 3. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28698547), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu, Rt

Species Glossary

Reactivity: Hu, Rt

Applications:

,

Applications: ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	AF2886U-100UG
Alexa Fluor 405	AF2886V-100UG
Alexa Fluor 488	AF2886G-100UG
Alexa Fluor 532	AF2886X-100UG
Alexa Fluor 594	AF2886T-100UG
Alexa Fluor 647	AF2886R-100UG
Alexa Fluor 700	AF2886N-100UG
Alexa Fluor 750	AF2886S-100UG
Alexa Fluor Plus 405	AF2886AFP405-100UG
Alexa Fluor Plus 488	AF2886AFP488-100UG
Alexa Fluor Plus 555	AF2886AFP555-100UG
Alexa Fluor Plus 594	AF2886AFP594-100UG
Alexa Fluor Plus 647	AF2886AFP647-100UG
Alexa Fluor Plus 680	AF2886AFP680-100UG
Allophycocyanin	AF2886A
Biotin	AF2886B
CoraFluor 1	AF2886CL1
DyLight 350	AF2886D
DyLight 405	AF2886E
DyLight 488	AF2886K
DyLight 550	AF2886L
DyLight 594	AF2886M
DyLight 650	AF2886W
DyLight 680	AF2886Y
DyLight 755	AF2886Z
FITC	AF2886F
HRP	AF2886H
Janelia Fluor 525	AF2886JF525
Janelia Fluor 549	AF2886I
Janelia Fluor 646	AF2886J
Janelia Fluor 669	AF2886JF669
PE	AF2886P
PerCP	AF2886C
mFluor Violet 450 SE	AF2886MFV450
mFluor Violet 500 SE	AF2886MFV500
mFluor Violet 610 SE	AF2886MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF2886-SP

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NBP2-76454

HIF-2 alpha/EPAS1 Antibody (2444A) - BSA Free

(1 Review)

0.1 mg

NBP2-76454

Immunohistochemistry-Paraffin: HIF-2 alpha/EPAS1 Antibody (2444A) [NBP2-76454] - Analysis of an FFPE tissue section of the human renal carcinoma using 1:100 dilution of HIF-2 alpha antibody (NBP2-76454). The signal was developed using HRP-DAB method which followed counterstaining of the cells with hematoxylin. The antibody generated staining mainly in the nuclear with minor cytoplasmic staining.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

,

Applications: ,

Host:

Rabbit Monoclonal

Host: Rabbit Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP2-76454AF350
Alexa Fluor 405	NBP2-76454AF405
Alexa Fluor 488	NBP2-76454AF488
Alexa Fluor 532	NBP2-76454AF532
Alexa Fluor 594	NBP2-76454AF594
Alexa Fluor 647	NBP2-76454AF647
Alexa Fluor 700	NBP2-76454AF700
Alexa Fluor 750	NBP2-76454AF750
Alexa Fluor Plus 405	NBP2-76454AFP405
Alexa Fluor Plus 488	NBP2-76454AFP488
Alexa Fluor Plus 555	NBP2-76454AFP555
Alexa Fluor Plus 594	NBP2-76454AFP594
Alexa Fluor Plus 647	NBP2-76454AFP647
Alexa Fluor Plus 680	NBP2-76454AFP680
Allophycocyanin	NBP2-76454APC
Biotin	NBP2-76454B
CoraFluor 1	NBP2-76454CL1
DyLight 350	NBP2-76454UV
DyLight 405	NBP2-76454V
DyLight 488	NBP2-76454G
DyLight 550	NBP2-76454R
DyLight 594	NBP2-76454DL594
DyLight 650	NBP2-76454C
DyLight 680	NBP2-76454FR
DyLight 755	NBP2-76454IR
FITC	NBP2-76454F
HRP	NBP2-76454H
Janelia Fluor 525	NBP2-76454JF525
Janelia Fluor 549	NBP2-76454JF549
Janelia Fluor 646	NBP2-76454JF646
Janelia Fluor 669	NBP2-76454JF669
PE	NBP2-76454PE
PerCP	NBP2-76454PCP
mFluor Violet 450 SE	NBP2-76454MFV450
mFluor Violet 500 SE	NBP2-76454MFV500
mFluor Violet 610 SE	NBP2-76454MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP2-76454SS
Azide and BSA Free	NBP2-80767

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MAB2886

HIF-2 alpha/EPAS1 Antibody (318404)

(1 Publication)

100 μg

MAB2886

HAF007). A specific band was detected for HIF-2a/EPAS1 at approximately 110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox">

Western blot shows lysates of 786-O human renal cell adenocarcinoma cell line (HIF-2a positive), Caki-2 human clear cell carcinoma epithelial cell line (HIF-2a negative), and HeLa human cervical epithelial carcinoma cell line (HIF-2a negative). PVDF Membrane was probed with 1 µg/mL of Human HIF-2a/EPAS1 Monoclonal Antibody (Catalog # MAB2886) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # <a class=

HAF007). A specific band was detected for HIF-2a/EPAS1 at approximately 110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of 786-O human renal cell adenocarcinoma cell line (HIF-2a positive), Caki-2 human clear cell carcinoma epithelial cell line (HIF-2a negative), and HeLa human cervical epithelial carcinoma cell line (HIF-2a negative). PVDF Membrane was probed with 1 µg/mL of Human HIF-2a/EPAS1 Monoclonal Antibody (Catalog # MAB2886) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for HIF-2a/EPAS1 at approximately 110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

Applications:

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	FAB2886U-100UG
Alexa Fluor 405	FAB2886V-100UG
Alexa Fluor 488	FAB2886G-100UG
Alexa Fluor 532	FAB2886X-100UG
Alexa Fluor 594	FAB2886T-100UG
Alexa Fluor 647	FAB2886R-100UG
Alexa Fluor 700	FAB2886N-100UG
Alexa Fluor 750	FAB2886S-100UG
Alexa Fluor Plus 405	FAB2886AFP405-100UG
Alexa Fluor Plus 488	FAB2886AFP488-100UG
Alexa Fluor Plus 555	FAB2886AFP555-100UG
Alexa Fluor Plus 594	FAB2886AFP594-100UG
Alexa Fluor Plus 647	FAB2886AFP647-100UG
Alexa Fluor Plus 680	FAB2886AFP680-100UG
Allophycocyanin	FAB2886A
Biotin	FAB2886B
CoraFluor 1	FAB2886CL1
DyLight 350	FAB2886D
DyLight 405	FAB2886E
DyLight 488	FAB2886K
DyLight 550	FAB2886L
DyLight 594	FAB2886M
DyLight 650	FAB2886W
DyLight 680	FAB2886Y
DyLight 755	FAB2886Z
FITC	FAB2886F
HRP	FAB2886H
Janelia Fluor 525	FAB2886JF525
Janelia Fluor 549	FAB2886I
Janelia Fluor 646	FAB2886J
Janelia Fluor 669	FAB2886JF669
PE	FAB2886P
PerCP	FAB2886C
mFluor Violet 450 SE	FAB2886MFV450
mFluor Violet 500 SE	FAB2886MFV500
mFluor Violet 610 SE	FAB2886MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	MAB2886-SP

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NBP3-07075

HIF-2 alpha/EPAS1 Antibody (2597C) - BSA Free

0.1 mg

NBP3-07075

Reactivity:

Hu, Mu

Species Glossary

Reactivity: Hu, Mu

Applications:

Applications:

Host:

Rabbit Monoclonal

Host: Rabbit Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP3-07075AF350
Alexa Fluor 405	NBP3-07075AF405
Alexa Fluor 488	NBP3-07075AF488
Alexa Fluor 532	NBP3-07075AF532
Alexa Fluor 594	NBP3-07075AF594
Alexa Fluor 647	NBP3-07075AF647
Alexa Fluor 700	NBP3-07075AF700
Alexa Fluor 750	NBP3-07075AF750
Alexa Fluor Plus 405	NBP3-07075AFP405
Alexa Fluor Plus 488	NBP3-07075AFP488
Alexa Fluor Plus 555	NBP3-07075AFP555
Alexa Fluor Plus 594	NBP3-07075AFP594
Alexa Fluor Plus 647	NBP3-07075AFP647
Alexa Fluor Plus 680	NBP3-07075AFP680
Allophycocyanin	NBP3-07075APC
Biotin	NBP3-07075B
CoraFluor 1	NBP3-07075CL1
DyLight 350	NBP3-07075UV
DyLight 405	NBP3-07075V
DyLight 488	NBP3-07075G
DyLight 550	NBP3-07075R
DyLight 594	NBP3-07075DL594
DyLight 650	NBP3-07075C
DyLight 680	NBP3-07075FR
DyLight 755	NBP3-07075IR
FITC	NBP3-07075F
HRP	NBP3-07075H
Janelia Fluor 525	NBP3-07075JF525
Janelia Fluor 549	NBP3-07075JF549
Janelia Fluor 646	NBP3-07075JF646
Janelia Fluor 669	NBP3-07075JF669
PE	NBP3-07075PE
PerCP	NBP3-07075PCP
mFluor Violet 450 SE	NBP3-07075MFV450
mFluor Violet 500 SE	NBP3-07075MFV500
mFluor Violet 610 SE	NBP3-07075MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP3-07075SS

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NBL1-10286

HIF-2 alpha/EPAS1 Overexpression Lysate

(2 Publications)

0.1 mg

NBL1-10286

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

Applications:

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H00002034-P03

Recombinant Human HIF-2 alpha/EPAS1 GST (N-Term) Protein

(2 Publications)

2 μg

H00002034-P03

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

, , ,

Applications: , , ,

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NB100-122PEP

HIF-2 alpha/EPAS1 Antibody Blocking Peptide

0.1 mg

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

Applications:

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H00002034-Q01

Recombinant Human HIF-2 alpha/EPAS1 GST (N-Term) Protein

10 μg

H00002034-Q01

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

, , , ,

Applications: , , , ,

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

Category

Protein/Target

Applications

Reactivity

Citation

Antibody Validation

Reviews

Host

Available Conjugates

Format

Modification

Clonality

Clone

Images Type

Isotype

Preservative

Protein Or Peptide

Research Areas

Has Sample Size

Promotional Item

Species Glossary

EPAS1 Products By Gene Name

HIF-2 alpha/EPAS1 Antibody - BSA Free

HIF-2 alpha/EPAS1 Antibody (ep190b)

HIF-2 alpha/EPAS1 Antibody [Unconjugated]

HIF-2 alpha/EPAS1 Antibody Pack

HIF-2 alpha/EPAS1 Antibody

HIF-2 alpha/EPAS1 Antibody (2444A) - BSA Free

HIF-2 alpha/EPAS1 Antibody (318404)

HIF-2 alpha/EPAS1 Antibody (2597C) - BSA Free

HIF-2 alpha/EPAS1 Overexpression Lysate

Recombinant Human HIF-2 alpha/EPAS1 GST (N-Term) Protein

HIF-2 alpha/EPAS1 Antibody Blocking Peptide

Recombinant Human HIF-2 alpha/EPAS1 GST (N-Term) Protein