HIF-2 alpha/EPAS1 Antibody (ep190b)

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.

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. [PMID: 23785417] Image using the HRP form of this antibody.

Immunofluorescence: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Analysis of HIF-2 alpha in human endometrium using HIF-2 alpha/EPAS1 Antibody (ep190b). Donkey anti-mouse Alexa Fluor 488 secondary antibody was used. Image from verified customer review.

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.

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. [PMID: 23785417] Image using the HRP form of this antibody.

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 (NB100-132) at 1 - 2 ug/ml.

Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [NB100-132] - Western blot analysis and quantification of HIF-2alpha 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 licence.

• Reactivity: Hu, Mu, Rt, Ha
• Applications: WB, Simple Western, ChIP, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, In vivo, IP, GS, ICC/IF
• Clone: ep190b
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

HIF-2 alpha/EPAS1 Antibody (ep190b) Summary

• Immunogen: The immunogen recognized by this HIF-2 alpha/EPAS1 Antibody (ep190b) maps to a region between amino acids 535-631. [UniProt# Q99814]
• Specificity: This HIF-2 alpha/EPAS1 Antibody (ep190b) is specific for HIF-2 alpha/EPAS1, and does not cross-react with HIF-1 alpha.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: EPAS1
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1-2 ug/ml
  • Simple Western 1:100
  • Chromatin Immunoprecipitation
  • ELISA 1:100-1:2000
  • Flow Cytometry 1:400
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry 1:150-1:300
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:150-1:300
  • Immunoprecipitation
  • In vivo assay
  • Gel Supershift Assay
  • Immunofluorescence
• Application Notes: This HIF-2 alpha (ep190b) antibody is useful for ELISA, Flow Cytometry, Immunohistochemistry on paraffin-embedded sections and Western Blot. In WB, it recognizes a band at approx. 118 kDa representing HIF-2 alpha. Use in Gel supper shift assay reported in scientific literature (PMID: 17404621). Use in Immunohistochemistry-Frozen reported in scientific literature (PMID: 24973414). Use in vivo reported in scientific literature (PMID: 23857308).
  
  In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
  The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Theoretical MW: 96.5 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  Human Kidney Whole Tissue Lysate (Adult Whole Normal)
  HIF-2 alpha/EPAS1 Overexpression Lysate
  HIF-2 alpha/EPAS1 Knockout HeLa Cell Lysate

Reactivity Notes

Ability to use HIF-2 alpha/EPAS1 Antibody (ep190b) in mouse is mixed with some positive and some negative results.

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: PBS with 1% BSA
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for HIF-2 alpha/EPAS1 Antibody (ep190b)

• Basic-helix-loop-helix-PAS protein MOP2
• BHLHE73
• Class E basic helix-loop-helix protein 73
• ECYT4
• endothelial PAS domain protein 1
• endothelial PAS domain-containing protein 1
• ep190b
• EPAS1
• EPAS-1
• HIF 2A
• HIF-1-alpha-like factor
• HIF-1alpha-like factor
• HIF2 alpha
• HIF-2 alpha
• hif2a angiogenesis
• HIF2A
• HIF-2-alpha
• HIF2-alpha
• HLF
• hypoxia-inducible factor 2 alpha
• Hypoxia-inducible factor 2-alpha
• Member of PAS protein 2
• MOP2
• PAS domain-containing protein 2
• PASD2

Background

Hypoxia contributes to the pathophysiology of human disease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension, congenital heart disease and chronic obstructive pulmonary disease (1). In cancer, and particularly solid tumors, hypoxia plays a critical role in the regulation of genes involved in stem cell renewal, epithelial to mesenchymal transition (EMT), metastasis and angiogenesis. In the tumor microenvironment (TME), hypoxia influences the properties and function of stromal cells (e.g., fibroblasts, endothelial and immune cells) and is a strong determinant of tumor progression (2,3).

HIF-1 or hypoxia inducible factor 1, is a transcription factor commonly referred to as a "master regulator of the hypoxic response" for its central role in the regulation of cellular adaptations to hypoxia. Similarly, HIF-2 alpha plays a role in cellular responses to hypoxia, but whereas HIF-1 alpha is ubiquitously expressed, HIF-2 alpha is predominantly expressed in the vascular endothelium at embryonic stages and after birth in select cells and tissue types (e.g., fibroblasts, hepatocytes and myocytes at 96kDa) (4). Following a similar mechanism to HIF-1 alpha, HIF-2 alpha is stabilized under hypoxic conditions by the formation of a heterodimer with an ARNT/HIF-1 beta subunit. Stable HIF-2 alpha-ARNT/HIF-1 beta heterodimers engage p300/CBP in the nucleus for binding to hypoxic response elements (HREs), inducing transcription, and thus regulation of genes (e.g., EPO, VEGFA). HIF-1 predominantly transactivates genes involved in glycolytic control and pro-apoptotic genes (e.g., LDHA and BNIP3), and HIF-2 regulates the expression of genes involved in invasion and stemness (e.g., MMP2, and OCT4). Common gene targets for HIF-1 and HIF-2 include VEGFA and GLUT1 (5).

The HIF-2 alpha subunit is rapidly targeted and degraded by the ubiquitin proteasome system under normoxic conditions. This process is mediated by oxygen-sensing enzymes, prolyl hydroxylase domain enzymes (PHDs), which catalyze the hydroxylation of key proline residues (Pro-405 and Pro-531) within the oxygen-dependent degradation domain of HIF-2 alpha (5). Once hydroxylated, HIF-2 alpha binds the von Hippel-Lindau tumor suppressor protein (pVHL) for subsequent ubiquitination and proteasomal degradation (5,6).

References

1. Semenza, G. L., Agani, F., Feldser, D., Iyer, N., Kotch, L., Laughner, E., & Yu, A. (2000). Hypoxia, HIF-1, and the pathophysiology of common human diseases. Advances in Experimental Medicine and Biology.

2.Muz, B., de la Puente, P., Azab, F., & Azab, A. K. (2015). The role of hypoxia in cancer progression, angiogenesis, metastasis, and resistance to therapy. Hypoxia. https://doi.org/10.2147/hp.s93413

3. Huang, Y., Lin, D., & Taniguchi, C. M. (2017). Hypoxia inducible factor (HIF) in the tumor microenvironment: friend or foe? Science China Life Sciences. https://doi.org/10.1007/s11427-017-9178-y

4. Hu, C.-J., Wang, L.-Y., Chodosh, L. A., Keith, B., & Simon, M. C. (2003). Differential Roles of Hypoxia-Inducible Factor 1 (HIF-1) and HIF-2 in Hypoxic Gene Regulation. Molecular and Cellular Biology. https://doi.org/10.1128/mcb.23.24.9361-9374.2003

5. Koh, M. Y., & Powis, G. (2012). Passing the baton: The HIF switch. Trends in Biochemical Sciences. https://doi.org/10.1016/j.tibs.2012.06.004

6. Koyasu, S., Kobayashi, M., Goto, Y., Hiraoka, M., & Harada, H. (2018). Regulatory mechanisms of hypoxia-inducible factor 1 activity: Two decades of knowledge. Cancer Science. https://doi.org/10.1111/cas.13483

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for HIF-2 alpha/EPAS1 Antibody (NB100-132)(166)

Publications using NB100-132

• Bacci L, Aiello A, Ripoli C et al. H19-Dependent Transcriptional Regulation of beta 3 and beta 4 Integrins Upon Estrogen and Hypoxia Favors Metastatic Potential in Prostate Cancer Int J Mol Sci Aug 17 2019 [PMID: 31426484] (ChIP, WB)
• Tang K, Yu Y, Zhu L et al. Hypoxia-reprogrammed tricarboxylic acid cycle promotes the growth of human breast tumorigenic cells Oncogene Aug 13 2019 [PMID: 31409901] (WB, Mouse)
• Das B, Pal B, Bhuyan R et al. MYC Regulates the HIF2 alpha Stemness Pathway via Nanog and Sox2 to Maintain Self-Renewal in Cancer Stem Cells versus Non-Stem Cancer Cells Cancer Res. Jul 2 2019 [PMID: 31266772] (Flow, IHC, WB, Mouse, Human)
• Knowles HJ The Adenosine A2B Receptor Drives Osteoclast-Mediated Bone Resorption in Hypoxic Microenvironments Cells 2019 Jun 21 [PMID: 31234425] (WB, Human)
• Duan LJ, Fong GH. Developmental vascular pruning in neonatal mouse retinas is programmed in the astrocytic oxygen sensing mechanism Development Mar 25 2019 [PMID: 30910827] (WB, Mouse)
• Sasagawa T, Nagamatsu T, Morita K et al. HIF-2a, but not HIF-1a, mediates hypoxia-induced up-regulation of Flt-1 gene expression in placental trophoblasts. Sci Rep. Nov 26 2018 [PMID: 30478339] (WB, Human)
• Krueger K, Catanese L, Sciesielski LK et al. Deletion of an intronic HIF-2a binding site suppresses hypoxia-induced WT1 expression. Biochim Biophys Acta Gene Regul Mech. Nov 20 2018 [PMID: 30468780] (ChIP, Human)
• Liu N, Luo J, Kuang D et al. Lactate inhibits ATP6V0d2 expression in tumor-associated macrophages to promote HIF-2a-mediated tumor progression. J. Clin. Invest. Nov 15 2018 [PMID: 30431439] (Mouse)
• Han JS, Lee JH, Kong J et al. Hypoxia Restrains Lipid Utilization via Protein Kinase A and ATGL Downregulation through Hypoxia Inducible Factor. Mol. Cell. Biol. Nov 5 2018 [PMID: 30397073] (WB, Mouse)
• Moreno Roig E, Yaromina A, Houben R et al. Prognostic Role of Hypoxia-Inducible Factor-2alpha Tumor Cell Expression in Cancer Patients: A Meta-Analysis Front Oncol Jun 11 2018 [PMID: 29942795] (IHC-P, Human)
• Lin Z, Liu F, Shi P et al. Fatty acid oxidation promotes reprogramming by enhancing oxidative phosphorylation and inhibiting protein kinase C. Stem Cell Res Ther. Feb 26 2018 [PMID: 29482657] (WB, Mouse)
• Liu Y, Tan X, Liu W, Chen X. Follistatin-like protein 1 plays a tumor suppressor role in clear-cell renal cell carcinoma. Chin J Cancer. 2018 Jan 22 [PMID: 29357946] (IHC-P, Human)
• Chen R, Xu M, Nagati J, Garcia JA. Coordinate regulation of stress signaling and epigenetic events by Acss2 and HIF-2 in cancer cells. PLoS ONE 2017 Dec 27 [PMID: 29281714] (WB, IP, ChIP, Human)
• Carew JS, Espitia CM, Zhao W et al. Oncolytic reovirus inhibits angiogenesis through induction of CXCL10/IP-10 and abrogation of HIF activity in soft tissue sarcomas Oncotarget. 2017 Oct 17 [PMID: 29156834] (WB)
• Sadiku P, Willson JA, Dickinson RS et al. Prolyl hydroxylase 2 inactivation enhances glycogen storage and promotes excessive neutrophilic responses J. Clin. Invest. 2017 Aug 14 [PMID: 28805660] (IHC-P, Mouse)
• Padro M, Louie RJ, Lananna BV et al. Genome-independent hypoxic repression of estrogen receptor alpha in breast cancer cells. BMC Cancer. Mar 20 2017 [PMID: 28320353] (WB, ICC/IF, Human)
• Liu W, Chen H, Wong N et al. Pseudohypoxia induced by miR-126 deactivation promotes migration and therapeutic resistance in renal cell carcinoma. Cancer Lett. May 28 2017 [PMID: 28257806] (WB, Human)
• Shiraishi A, Tachi K, Essid N et al. Hypoxia promotes the phenotypic change of aldehyde dehydrogenase activity of breast cancer stem cells. Cancer Sci. Mar 1 2017 [PMID: 28012234] (WB, Human)
• Endo H, Okami J, Okuyama H et al. The induction of MIG6 under hypoxic conditions is critical for dormancy in primary cultured lung cancer cells with activating EGFR mutations. Oncogene. Nov 28 2016 [PMID: 27893711] (WB, Human)
• Leszczynska KB, Dobrynin G, Leslie RE et al. Preclinical testing of an Atr inhibitor demonstrates improved response to standard therapies for esophageal cancer. Radiother Oncol. Nov 1 2016 [PMID: 27839769] (WB, Human)
• Kwak JH, Lee NH, Lee HY et al. HIF2a/EFEMP1 cascade mediates hypoxic effects on breast cancer stem cell hierarchy Oncotarget Jul 12 2016 [PMID: 27270657] (WB, Mouse)
• Szendroi A, Szasz AM, Kardos M et al. Opposite prognostic roles of HIF1alpha and HIF2alpha expressions in bone metastatic clear cell renal cell cancer. Oncotarget. 2016 May 27 [PMID: 27244898] (IHC)
  
  Details:
  This citation used the HRP version of this antibody.
• Nusken E, Wohlfarth M, Lippach G et al. Reduced Perinatal Leptin Availability May Contribute to Adverse Metabolic Programming in a Rat Model of Uteroplacental Insufficiency. Endocrinology. 2016 May [PMID: 27007072]
  
  Details:
  This citation used the HRP version of this antibody.
• Espana-Agusti J, Zou X, Wong K et al. Generation and Characterisation of a Pax8-CreERT2 Transgenic Line and a Slc22a6-CreERT2 Knock-In Line for Inducible and Specific Genetic Manipulation of Renal Tubular Epithelial Cells. PLoS One 2016 Feb 11 [PMID: 26866916]
• Kim Y, Nam HJ, Lee J et al. Methylation-dependent regulation of HIF-1alpha stability restricts retinal and tumour angiogenesis. Nat Commun 2016 Jan 13 [PMID: 26757928] (WB)
• Lee SB, Frattini V, Bansal M et al. An ID2-dependent mechanism for VHL inactivation in cancer. Nature. 2016 Jan 14 [PMID: 26735018] (IHC-P, Human)
• Su J, Li Z, Cui S et al. The Local HIF-2alpha/EPO Pathway in the Bone Marrow is Associated with Excessive Erythrocytosis and the Increase in Bone Marrow Microvessel Density in Chronic Mountain Sickness High Alt Med Biol [PMID: 26625252] (Human)
  
  Details:
  Used the DyLight 488 form of this antibody.
• Seidl CI, Martinez-Sanchez A, Murphy CL. Derepression of miRNA-138 contributes to loss of the human articular chondrocyte phenotype. Arthritis & rheumatology 2015 Sep 11 [PMID: 26359943] (WB, Human)
• Espana-Agusti J, Tuveson DA, Adams DJ, Matakidou A. A minimally invasive, lentiviral based method for the rapid and sustained genetic manipulation of renal tubules. Sci Rep 2015 [PMID: 26046460] (IHC, Mouse)
• Her YF, Nelson-Holte M, Maher LJ. Oxygen concentration controls epigenetic effects in models of familial paraganglioma. PLoS ONE. 2015 May 19 [PMID: 25985299] (WB, IHC-P, Human, Mouse)
• Pahlman S, Lund LR, Jogi A. Differential HIF-1alpha and HIF-2alpha Expression in Mammary Epithelial Cells during Fat Pad Invasion, Lactation, and Involution. PLoS ONE. 2015 May 09 [PMID: 25955753] (IHC-P, Mouse)
• Chen R, Xu M, Nagati JS et al. The Acetate/ACSS2 Switch Regulates HIF-2 Stress Signaling in the Tumor Cell Microenvironment PLoS ONE. 2015 Feb 18 [PMID: 25689462] (WB, IP, Human)
• Chen T, Zhou G, Zhou Q et al. Loss of miR-17~92 in Smooth Muscle Cells Attenuates Experimental Pulmonary Hypertension via Induction of PDLIM5 Am. J. Respir. Crit. Care Med. 2015 Feb 03 [PMID: 25647182] (WB, Human)
• Niemi Henna, Honkonen Krista, Korpisalo Petra et al. HIF-1alpha and HIF-2alpha induce angiogenesis and improve muscle energy recovery. Eur J Clin Invest. 2014 Oct 01 [PMID: 25208310]
• Xu M, Nagati JS, Xie J et al. An acetate switch regulates stress erythropoiesis. Nat. Med. 2014 Aug 10 [PMID: 25108527] (WB, ChIP, IP, Human)
• Motzer Robert J, Hutson Thomas E, Hudes Gary R et al. Investigation of novel circulating proteins, germ line single-nucleotide polymorphisms, and molecular tumor markers as potential efficacy biomarkers of first-line sunitinib therapy for advanced renal cell carcinoma. Cancer Chemother Pharmacol. 2014 Aug 07 [PMID: 25100134] (Human)
• Evans CE, Wadoodi A, Humphries J et al. Local accumulation of hypoxia-inducible factor 2 alpha during venous thrombus resolution. Thromb. Res. 2014 Jul 23 [PMID: 25087887] (IHC-P, Mouse)
  
  Details:
  HIF2a/HIF-2 alpha antibody used for IHC-P in venous thrombus induced in the inferior vena cava /IVC of male BALB/C mice treated or not with HIF agonist L-mimosine (Fig 1, 2). Antigen retrieval - pressure cooking sections in citrate buffer (0.3% sodium citrate, pH 6).
• Wong Waihay J, Richardson Theresa, Seykora John T et al. Hypoxia-inducible factors regulate filaggrin expression and epidermal barrier function. J Invest Dermatol. 2014 Jul 07 [PMID: 24999590] (Mouse)
• Peng YJ, Yuan G, Khan S et al. Regulation of Hypoxia Inducible Factor-alpha Isoforms and Redox State By Carotid Body Neural Activity in Rats. J. Physiol. (Lond.). 2014 Jun 27 [PMID: 24973414] (IHC-Fr, WB, Rat)
• Carrera S, Senra J, Acosta MI et al. The Role of the HIF-1alpha Transcription Factor in Increased Cell Division at Physiological Oxygen Tensions. PLoS ONE. 2014 May 19 [PMID: 24835245] (WB, Human)
• Toyoda H, Hirayama J, Sugimoto Y et al. Polycythemia and Paraganglioma With a Novel Somatic HIF2A Mutation in a Male. Pediatrics. 2014 May 12 [PMID: 24819565] (IHC-P, Human)
• Gregg JL, Turner RM, Chang G et al. NADPH oxidase NOX4 supports renal tumorigenesis by promoting the expression and nuclear accumulation of HIF2alpha. Cancer Res. 2014 Apr 22 [PMID: 24755467] (ICC/IF, IHC-P, WB, Human)
• Raspaglio G, Petrillo M, Martinelli E et al. Sox9 and Hif-2a regulate TUBB3 gene expression and affect ovarian cancer aggressiveness. Gene 3/21/2014 [PMID: 24661907] (ChIP, WB, Human)
• Duan LJ, Takeda K, Fong GH. Hematological, Hepatic, and Retinal Phenotypes in Mice Deficient for Prolyl Hydroxylase Domain (PHD) Proteins in the Liver. Am. J. Pathol. 2014 Feb 19 [PMID: 24508125] (WB, Mouse)
• Duan LJ, Takeda K, Fong GH. Hypoxia inducible factor-2 alpha regulates the development of retinal astrocytic network by maintaining adequate supply of astrocyte progenitors. PLoS ONE 2014 Jan 29 [PMID: 24475033] (WB, Mouse)
• Ellinghaus P, Heisler I, Unterschemmann K et al. BAY 87-2243, a highly potent and selective inhibitor of hypoxia-induced gene activation has antitumor activities by inhibition of mitochondrial complex I. Cancer Med 2013 Oct [PMID: 24403227] (WB)
• Anderson ER, Taylor M, Xue X et al. Intestinal HIF2alpha promotes tissue-iron accumulation in disorders of iron overload with anemia. Proc Natl Acad Sci U S A. 2013 Dec 10 [PMID: 24282296] (WB, Mouse)
• Zhang T, Niu X, Liao L et al. The Contributions of HIF-Target Genes to Tumor Growth in RCC. PLoS One. 2013 Nov 18 [PMID: 24260413] (WB, Human)
• Tsang WP, Shu Y, Kwok PL et al. CD146(+) Human Umbilical Cord Perivascular Cells Maintain Stemness under Hypoxia and as a Cell Source for Skeletal Regeneration. PLoS One. 2013 Oct 18 [PMID: 24204598] (WB, Human)
• Thompson AA, Elks PM, Marriott HM et al. Hypoxia-inducible factor 2alpha regulates key neutrophil functions in humans, mice and zebrafish. Blood. 2013 Nov 6 [PMID: 24196071] (WB, IHC, Human, Mouse)
• Klotzsche-von Ameln A, Prade I, Grosser M et al. Prolyl hydroxylase-4 overexpression stimulates tumor angiogenesis in mouse osteosarcoma via transforming growth factor-alpha. Mol Cancer Re. 2013 Sep 18 [PMID: 24048703] (WB, Human)
• Yeh IJ, Ogba N, Bensigner H et al. HEXIM1 downregulates Hypoxia Inducible Factor-1 alpha protein stability. Biochem J. 2013 Sep 10 [PMID: 24015760] (WB, Human)
• David JM, Owens TA, Barwe SP, Rajasekaran AK. Gramicidin a induces metabolic dysfunction and energy depletion leading to cell death in renal cell carcinoma cells. Mol Cancer Ther. 2013 Nov [PMID: 24006494]
• Bruells CS, Maes K, Rossaint R et al. Prolonged Mechanical Ventilation Alters the Expression Pattern of Angio-neogenetic Factors in a Pre-Clinical Rat Model. PLoS One. 2013 Aug 8 [PMID: 23950950] (WB, Rat)
• Park YK, Park B, Lee SY et al. Hypoxia-Inducible Factor-2alpha-Dependent Hypoxic Induction of Wnt10b Expression in Adipogenic Cells. J Biol Chem 2013 Jul 30 [PMID: 23900840] (WB, Mouse)
• Gottfried E, Lang SA, Renner K et al. New Aspects of an Old Drug - Diclofenac Targets MYC and Glucose Metabolism in Tumor Cells. PLoS One 2013 Jul 9 [PMID: 23874405] (WB, Human)
• Nakajima T, Nakajima E, Shearer TR, Azuma M. Concerted inhibition of HIF-1alpha and -2alpha expression markedly suppresses angiogenesis in cultured RPE cells. Mol Cell Biochem 2013 Jul 20 [PMID: 23873332] (WB, Human)
• Jamil S, Cedervall J, Hultman I et al. Neuroblastoma cells injected into experimental mature teratoma reveal a tropism for embryonic loose mesenchyme. Int J Oncol 2013 Sep [PMID: 23857308]
• Adamski J, Price A, Dive C, Makin G. Hypoxia-Induced Cytotoxic Drug Resistance in Osteosarcoma Is Independent of HIF-1Alpha. PLoS One 2013 Jun 13 [PMID: 23785417] (WB, Human)
• Nakayama K. CREB and NF-kB are activated during prolonged hypoxia and cooperatively regulate the induction of matrix metalloproteinase MMP1. J Biol Chem 2013 Jun 17 [PMID: 23775082] (WB, Human)
• Fukui N, Kageyama Y, Higashi Y et al. Development of a novel interferon-alpha2b gene construct with a repetitive hypoxia-inducible factor binding site and its suppressive effects on human renal cell carcinoma cell lines in vitro. Int J Clin Oncol 2013 Jun 6 [PMID: 23739923] (WB, Human)
• Lee WH, Lee JM, Lim C, Kim S, Kim SG et al. Structural requirements within protoporphyrin IX in the inhibition of heat shock protein 90. Chem Biol Interact 2013 Apr 24 [PMID: 23624237] (WB, Human)
• Kucharzewska P, Christianson HC, Welch JE et al. Exosomes reflect the hypoxic status of glioma cells and mediate hypoxia-dependent activation of vascular cells during tumor development. Proc Natl Acad Sci U S A 2013 Apr 30 [PMID: 23589885] (WB, Human)
• Onnis B, Fer N, Rapisarda A et al. Autocrine production of IL-11 mediates tumorigenicity in hypoxic cancer cells J Clin Invest. 2013 Apr [PMID: 23549086] (WB, Human)
• Su J, Li Z, Cui S et al. The Local HIF-2alpha/EPO Pathway in the Bone Marrow is Associated with Excessive Erythrocytosis and the Increase in Bone Marrow Microvessel Density in Chronic Mountain Sickness High Alt Med Biol (Human)
  
  Details:
  This publication used the DyLight 488 conjugated form of this antibody (Cat# NB100-132G).
• Scheuermann TH, Li Q, Ma HW et al. Allosteric inhibition of hypoxia inducible factor-2 with small molecules. Nat Chem Biol 2013 Apr [PMID: 23434853] (ChIP, Human)
• Huang C, Hales BF. Effects of exposure to a DNA damaging agent on the hypoxia inducible factors in organogenesis stage mouse limbs PLoS One 2012 [PMID: 23251655] (WB, IHC, Mouse)
• Sun HX, Xu Y, Yang XR et al. Hypoxia inducible factor 2 alpha inhibits hepatocellular carcinoma growth through the transcription factor dimerization partner 3/ E2F transcription factor 1-dependent apoptotic pathway. Hepatology 2012 Dec 5 [PMID: 23212661] (ICC/IF, Human)
• Macaulay IC, Thon JN, Tijssen MR et al. Canonical Wnt signaling in megakaryocytes regulates proplatelet formation Blood 2012 Nov 16 [PMID: 23160460]
• Havasi A, Haegele JA, Gall JM et al. Histone Acetyl Transferase (HAT) HBO1 and JADE1 in Epithelial Cell Regeneration Am J Pathol 2012 Nov 14 [PMID: 23159946]
• Persano L, Pistollato F, Rampazzo E et al. BMP2 sensitizes glioblastoma stem-like cells to Temozolomide by affecting HIF-1 alpha stability and MGMT expression Cell Death Dis 2012 Oct 18 [PMID: 23076220] (WB, ChIP, Human)
• Demachi-Okamura A, Torikai H, Akatsuka Y et al. Autophagy Creates a CTL Epitope That Mimics Tumor-Associated Antigens PLoS One 2012 [PMID: 23071732] (WB, Human)
• Xue X, Shah YM. Hypoxia-inducible factor-2 alpha is essential in activating the COX2/mPGES-1/PGE2 signaling axis in colon cancer Carcinogenesis 2012 Oct 5 [PMID: 23042097] (ChIP, Mouse)
• Vaapil M, Helczynska K, Villadsen R et al. Hypoxic Conditions Induce a Cancer-Like Phenotype in Human Breast Epithelial Cells PLoS One 2012 [PMID: 23029547] (IHC, WB, Human)
• Thomas S, Harding MA, Smith SC et al. CD24 Is an Effector of HIF-1-Driven Primary Tumor Growth and Metastasis Cancer Res 2012 Oct 22 [PMID: 22926560] (WB, Human)
• Muratsu-Ikeda S, Nangaku M, Ikeda Y et al. Downregulation of miR-205 Modulates Cell Susceptibility to Oxidative and Endoplasmic Reticulum Stresses in Renal Tubular Cells plos One 2012 [PMID: 22859986] (WB, Human)
• Mihaylova S, Killian A, Mayer K et al. Effects of anti-inflammatory vagus nerve stimulation on the cerebral microcirculation in endotoxinemic rats J Neuroinflammation 2012 Jul 25 [PMID: 22830560] (WB, Rat)
• Chen R, Xu M, Hogg RT et al. The Acetylase/Deacetylase Couple CREB-binding Protein/Sirtuin 1 Controls Hypoxia-inducible Factor 2 Signaling J Biol Chem 2012 Aug 31 [PMID: 22807441] (IP, Human)
• Kiss J, Mollenhauer M, Walmsley SR et al. Loss of the Oxygen Sensor PHD3 Enhances the Innate Immune Response to Abdominal Sepsis J Immunol 2012 Aug 15 [PMID: 22786772] (WB, Mouse)
• Lim CS, Kiriakidis S, Paleolog EM, Davies AH. The effects of doxycycline and micronized purified flavonoid fraction on human vein wall remodeling are not hypoxia-inducible factor pathway-dependent J Vasc Surg 2012 Jun 5 [PMID: 22677007] (WB, Human)
• El Naggar A, Clarkson P, Zhang F et al. Expression and stability of hypoxia inducible factor 1alpha in osteosarcoma Pediatr Blood Cancer 2012 Dec 15 [PMID: 22544557] (WB, Human)
• Jeong CW, Yoon CY, Jeong SJ, Hong SK, Byun SS, Kwak C, Lee SE. The Role of Hypoxia-Inducible Factor-1? and -2? in Androgen Insensitive Prostate Cancer Cells. Urol Oncol. 2012 Apr 24. [PMID: 22537539] (WB, Human)
• Schodel J, Bardella C, Sciesielski LK et al. Common genetic variants at the 11q13.3 renal cancer susceptibility locus influence binding of HIF to an enhancer of cyclin D1 expression Nat Genet 2012 Mar 11 [PMID: 22406645] (WB, Human)
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  Details:
  Using the HRP conjugated version of NB100-132, catalog number NB100-132H.
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• Rankin EB, Biju MP, Liu Q et al. Hypoxia-inducible factor-2 (HIF-2) regulates hepatic erythropoietin in vivo. J Clin Invest 2007 Apr [PMID: 17404621] (EMSA, Mouse)
• Lau KW, Tian YM, Raval RR et al. Target gene selectivity of hypoxia-inducible factor-alpha in renal cancer cells is conveyed by post-DNA-binding mechanisms. Br J Cancer 2007 Apr 23 [PMID: 17387348] (WB, Human)
• Harvey, A J et al. Regulation of gene expression in bovine blastocysts in response to oxygen and the iron chelator desferrioxamine. Biol Reprod;77(1):93-101. 2007 Jul. [PMID: 17329595] (ICC/IF, Human)
• Bernhardt WM, Wiesener MS, Weidemann A et al. Involvement of hypoxia-inducible transcription factors in polycystic kidney disease. Am J Pathol;170(3):830-42. 2007 Mar. [PMID: 17322369] (IHC-P, Rat)
• Kyriacou HM, Steen KE, Raza A et al. In vitro inhibition of Plasmodium falciparum rosette formation by Curdlan sulfate. Antimicrob Agents Chemother 2007 Apr [PMID: 17283200] (IHC)
• Yan Q, Bartz S, Mao M, Li L, Kaelin WG Jr. The hypoxia-inducible factor 2alpha N-terminal and C-terminal transactivation domains cooperate to promote renal tumorigenesis in vivo. Mol Cell Biol;27(6):2092-102. 2007 Mar. [PMID: 17220275]
• Koizume S, Jin MS, Miyagi E et al. Activation of cancer cell migration and invasion by ectopic synthesis of coagulation factor VII. Cancer Res;66(19):9453-60. 2006 Oct 1. [PMID: 17018600] (ChIP)
• Khan Z, Michalopoulos GK, Stolz DB. Peroxisomal localization of hypoxia-inducible factors and hypoxia-inducible factor regulatory hydroxylases in primary rat hepatocytes exposed to hypoxia-reoxygenation. Am J Pathol 2006 Oct [PMID: 17003483] (Rat)
• Carroll, P A et al. Latent Kaposi's sarcoma-associated herpesvirus infection of endothelial cells activates hypoxia-induced factors. J Virol;80(21):10802-12. 2006 Nov. [PMID: 16956952] (IHC-P, Human)
• Shinojima, Toshiaki et al. Cancer Biology: Renal cancer cells lacking hypoxia inducible factor (HIF)-1a expression maintain vascular endothelial growth factor expression through HIF-2a. Carcinogenesis;28(3):529-36. 2007 Mar. [PMID: 16920734] (WB)
• Zheng X, Ruas JL, Cao R, Salomons FA, Cao Y, Poellinger L, Pereira T. Cell-type-specific regulation of degradation of hypoxia-inducible factor 1 alpha: role of subcellular compartmentalization. Mol Cell Biol;26(12):4628-41. 2006 Jun. [PMID: 16738327]
• Kim CM, Vocke C, Torres-Cabala C et al. Expression of hypoxia inducible factor-1alpha and 2alpha in genetically distinct early renal cortical tumors. J Urol 2006 May [PMID: 16600797] (Human)
• Nakamura E, Abreu-e-Lima P, Awakura Y, Inoue T, Kamoto T, Ogawa O, Kotani H, Manabe T, Zhang GJ, Kondo K, Nose V, Kaelin WG Jr. Clusterin is a secreted marker for a hypoxia-inducible factor-independent function of the von Hippel-Lindau tumor suppressor protein. Am J Pathol;168(2):574-84. 2006 Feb. [PMID: 16436671]
• Bastian Eul, Frank Rose,Stefanie Krick. Impact of HIF-1#945; HIF-2#945; on proliferation migration of human pulmonary artery fibroblasts in hypoxia. The FASEB Journal express article 10.1096/fj.05-4104fje. [PMID: 16263938]
• Li, L et al. Evaluating Hypoxia-Inducible Factors-1alpha as a Cancer Therapeutic Target via Inducible RNA Interference In vivo. American Association for Cancer Research 65, 7249-7258. 2005 [PMID: 16103076] (WB, IHC)
• Isaacs JS, Jung YJ, Mole DR et al. HIF overexpression correlates with biallelic loss of fumarate hydratase in renal cancer: novel role of fumarate in regulation of HIF stability. Cancer Cell 2005 Aug [PMID: 16098467] (ELISA, Human)
• Bilton R, Mazure N, Trottier E et al. Arrest-defective-1 protein, an acetyltransferase, does not alter stability of hypoxia-inducible factor (HIF)-1alpha and is not induced by hypoxia or HIF. J Biol Chem 2005 Sep 02 [PMID: 15994306] (WB, Human)
• Yoshimura H, Dhar DK, Kohno H, Kubota H, Fujii T, Ueda S, Kinugasa S, Tachibana M, Nagasue N. Prognostic impact of hypoxia-inducible factors 1alpha and 2alpha in colorectal cancer patients: correlation with tumor angiogenesis and cyclooxygenase-2 expression. Clin Cancer Res;10(24):8554-60. 2004 Dec 15. [PMID: 15623639]
• Rathmell WK, Hickey MM, Bezman NA, Chmielecki CA, Carraway NC, Simon MC. In vitro and in vivo models analyzing von Hippel-Lindau disease-specific mutations. Cancer Res;64(23):8595-603. 2004 Dec 1. [PMID: 15574766]
• Grabmaier, K et al. Strict regulation of CAIXG250/MN by HIF-1 alpha in clear cell renal cell carcinoma. Oncogene;23(33):5624-31. 2004 Jul 22. [PMID: 15184875] (Human)
• Mekhail, K et al. HIF activation by pH-dependent nucleolar sequestration of VHL. Nature Cell Biology. 6: 642-647. 2004 [PMID: 15181450]
• Harvey AJ, Kind KL, Pantaleon M, Armstrong DT, Thompson JG. Oxygen-regulated gene expression in bovine blastocysts. Biol Reprod;71(4):1108-19. 2004 Oct. [PMID: 15163614]
• Koike, T et al. Hypoxia induces adhesion molecules on cancer cells: A missing link between Warburg effect induction of selectin-lig carbohydrates. PNAS. 101:8132-8137. 2004 [PMID: 15141079]
• Tissot van Patot MC, Bendrick-Peart J, Beckey VE, Serkova N, Zwerdlinger L. Greater vascularity, lowered HIF-1/DNA binding, and elevated GSH as markers of adaptation to in vivo chronic hypoxia. Am J Physiol Lung Cell Mol Physiol;287(3):L525-32. 2004 Sep. [PMID: 15132953]
• Pan Y, Oprysko PR, Asham AM, Koch CJ, Simon MC. p53 cannot be induced by hypoxia alone but responds to the hypoxic microenvironment. Oncogene;23(29):4975-83. 2004 Jun 24. [PMID: 15107830]
• Zimmer, M et al. Inhibition of Hypoxia-Inducible Factor Is Sufficient for Growth Supression of VHL-/- Tumors. Molecular Cancer Research 2: 89-95. 2004 [PMID: 14985465] (WB)
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Reviews for HIF-2 alpha/EPAS1 Antibody (NB100-132) (7)

Average Rating: 4.3

(Based on 7 reviews)

We have 7 reviews tested in 2 species: Human, Mouse.

reviewed by: Anonymous IF Human 02/05/2016

Summary

• Application: Immunofluorescence
• Sample Tested: Human endometrium
• Species: Human
• Lot: lot J-3

Blocking

• Blocking Details: 10% normal serum

Primary Anitbody

• Dilution Ratio: 1:250 in PBS, overnight incubation at 4 c

Secondary Antibody

• Secondary Description: alexa fluor 488 donkey anti mouse
• Secondary Manufacturer Cat#: Jackson Immunolab
• Secondary Concentration: 1:500

Details

• Detection Notes: negative control without primary antibody
• Fixation Details: antigen retrieval
• Wash Description: wash 3X PBS

reviewed by: Darren Richard WB Mouse 07/24/2015

Summary

• Application: Western Blot
• Sample Tested: Endothelial
• Species: Mouse

Blocking

• Blocking Details: TBS-T, Milk

Primary Anitbody

• Dilution Ratio: 1:1000

Secondary Antibody

• Secondary Description: Goat anti-mouse IgG, HRP
• Secondary Manufacturer Cat#: Promega
• Secondary Concentration: 1:2000

Details

• Detection Notes: Homemade ECL

Comments

• Comments: A good choice for Western blots.

reviewed by: Anonymous Mouse 02/04/2015

Summary

• Species: Mouse
• File: View PDF

reviewed by: Anonymous WB Human 12/12/2014

Summary

• Application: Western Blot
• Sample Tested: See PMID 23549086
• Species: Human

Blocking

• Blocking Details: See PMID 23549086

Details

• Detection Notes: See PMID 23549086

Comments

• Comments: Published in PMID:23549086.

reviewed by: Salvador Macip WB Human 08/11/2014

Summary

• Application: Western Blot
• Species: Human

reviewed by: Anonymous WB Mouse 07/28/2014

Summary

• Application: Western Blot
• Sample Tested: Mouse Kidney Tissue C57B5
• Species: Mouse
• Comments: I used frozen tissue

Blocking

• Blocking Details: 1 hr blocked in 10% bovine serum albumin in phosphate buffered saline (PBS) containing 0.1% (v/v) Tween-20 (PBS-T)

Primary Anitbody

• Dilution Ratio: 1mg/ml 4C over night

Secondary Antibody

• Secondary Description: anti rabbit
• Secondary Concentration: 1:15,000

Details

• Detection Notes: western blots were developed using supersignal west femto maximum sensitivity

Comments

• Comments: I used frozen tissue

reviewed by: Anonymous WB Human 12/14/2011

Summary

• Application: Western Blot
• Sample Tested: Human
• Species: Human

Blocking

• Blocking Details: Blocking Buffer: 1% Milk

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:500, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees Celsius

Secondary Antibody

• Secondary Description: Secondary Ab Dilution Ratio: 1:2000

Details

• Detection Notes: Detection Method: ECL

FAQs for HIF-2 alpha/EPAS1 Antibody (NB100-132). (Showing 1 - 2 of 2 FAQs).

1. Do you have a specific protocol for using NB100-132 for western blot and a recipe for preparing cell lysates to test this antibody?
   • Please find the specific protocol for NB100-132 here. This HIF antibody will require the standard hypoxic cell treatment.
2. I tried the antibody. By western blot on mouse T cells, I obtained a major non-specific band at 70KDa and a very weak band at 100KDa. Do you know what the 70KDa protein could be? However, I did this experiment on total cell lysates and I would like to redo this on nuclear extracts.
   • We have also encountered a band at 70kDa in our testing on whole cell lysates. In our experience, this band usually does not respond to hypoxic induction, is only present in whole cell and cytoplasmic fractions, and is seen in almost all HIF1 alpha products that we have tested. We currently do not know the identity of this band, but we think it could be a HIF1 alpha degradation product.

Bioinformatics

• Gene Symbol: EPAS1
• Entrez:
  • 2034(Human)
  • 13819(Mouse)
  • 29452(Rat)
• Uniprot:
  • Q99814()