HIF-2 alpha/EPAS1 Antibody [Unconjugated]

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.

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 # 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.

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 CoCl2for 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 # 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.

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.

• Reactivity: Hu, Mu, Rt
• Applications: WB, IHC
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated

HIF-2 alpha/EPAS1 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant mouse HIF-2 alpha /EPAS1
  Ser542-Thr874
  Accession # AAH57870
• Specificity: Detects human, mouse, and rat HIF-2 alpha /EPAS1 in Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: EPAS1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for HIF-2 alpha/EPAS1 Antibody [Unconjugated]

• 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
• 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

The hypoxia-inducible transcription factor 2 alpha (HIF-2 alpha ) is stabilized in hypoxic tissue and, similarly to HIF-1 alpha , complexes with Aryl hydrocarbon receptor nuclear translocator (ARNT). Both the HIF-1 and HIF-2 complexes bind hypoxia-response elements (HREs) in the promoters of many genes involved in adapting to an environment of insufficient oxygen or hypoxia. HIF-1 and HIF-2 do not appear completely redundant, although specific functions are only beginning to be elucidated. Hypoxic tissue environments occur in vascular and pulmonary diseases as well as cancer, which illustrates the potentially broad impact of gene regulation by both HIF-1 alpha and HIF-2 alpha .

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 (AF2997)(34)

Publications using AF2997

• Seidel, LM;Thudium, J;Smith, C;Sapehia, V;Sommer, N;Wujak, M;Weissmann, N;Seeger, W;Schermuly, RT;Novoyatleva, T; Death-associated protein kinase 1 prevents hypoxia-induced metabolic shift and pulmonary arterial smooth muscle cell proliferation in PAH Cellular signalling 2024-11-30 [PMID: 39622428] (Western Blot, Mouse, Rat)
• Wulf, S;Mizko, L;Herrmann, KH;Sánchez-Carbonell, M;Urbach, A;Lemke, C;Berndt, A;Loeffler, I;Wolf, G; Targeted Disruption of the MORG1 Gene in Mice Causes Embryonic Resorption in Early Phase of Development Biomolecules 2023-06-24 [PMID: 37509073] (IHC, Mouse)
• AM Mello, T Ngodup, Y Lee, KL Donahue, J Li, A Rao, ES Carpenter, HC Crawford, M Pasca di M, KE Lee Hypoxia promotes an inflammatory phenotype of fibroblasts in pancreatic cancer Oncogenesis, 2022-09-15;11(1):56. 2022-09-15 [PMID: 36109493] (Western Blot, Mouse)
• A Jatho, A Zieseniss, K Brechtel-C, J Guo, KO Böker, G Salinas, RH Wenger, DM Katschinsk The HIFalpha-Stabilizing Drug Roxadustat Increases the Number of Renal Epo-Producing Sca-1+ Cells Cells, 2022-02-21;11(4):. 2022-02-21 [PMID: 35203399] (Western Blot, Mouse)
• J Protze, S Naas, R Krüger, C Stöhr, A Kraus, S Grampp, M Wiesener, M Schiffer, A Hartmann, B Wullich, J Schödel The renal cancer risk allele at 14q24.2 activates a novel hypoxia-inducible transcription factor-binding enhancer of DPF3 expression The Journal of Biological Chemistry, 2022-02-08;0(0):101699. 2022-02-08 [PMID: 35148991] (Western Blot, Human)
• Magdalena Wujak, Christine Veith, Cheng-Yu Wu, Tessa Wilke, Zeki Ilker Kanbagli, Tatyana Novoyatleva et al. Adenylate Kinase 4—A Key Regulator of Proliferation and Metabolic Shift in Human Pulmonary Arterial Smooth Muscle Cells via Akt and HIF-1 alpha Signaling Pathways International Journal of Molecular Sciences 2021-09-26 [PMID: 34638712]
• S Kaluz, Q Zhang, Y Kuranaga, H Yang, S Osuka, D Bhattachar, NS Devi, J Mun, W Wang, R Zhang, MM Goodman, HE Grossnikla, EG Van Meir Targeting HIF-activated collagen prolyl 4-hydroxylase expression disrupts collagen deposition and blocks primary and metastatic uveal melanoma growth Oncogene, 2021-07-03;0(0):. 2021-07-03 [PMID: 34218269] (Western Blot, Human)
• Veli�a P, Cunha PP, Vojnovic N et al. Modified Hypoxia-Inducible Factor Expression in CD8(+) T Cells Increases Antitumor Efficacy Cancer Immunology Research 2021-04-01 [PMID: 33602720]
• W Yang, T Yu, X Huang, AJ Bilotta, L Xu, Y Lu, J Sun, F Pan, J Zhou, W Zhang, S Yao, CL Maynard, N Singh, SM Dann, Z Liu, Y Cong Intestinal microbiota-derived short-chain fatty acids regulation of immune cell IL-22 production and gut immunity Nat Commun, 2020-09-08;11(1):4457. 2020-09-08 [PMID: 32901017] (Western Blot, Mouse)
• N Burrows, RJM Bashford-R, VJ Bhute, A Peñalver, JR Ferdinand, BJ Stewart, JEG Smith, M Deobagkar-, G Giudice, TM Connor, A Inaba, L Bergamasch, S Smith, MGB Tran, E Petsalaki, PA Lyons, M Espeli, BJP Huntly, KGC Smith, RJ Cornall, MR Clatworthy, PH Maxwell Dynamic regulation of hypoxia-inducible factor-1&amp;alpha activity is essential for normal B cell development Nat. Immunol., 2020-08-31;0(0):. 8/31/2020 [PMID: 32868930]
• K Toriuchi, H Kakita, T Tamura, S Takeshita, Y Yamada, M Aoyama Prolonged astrocyte-derived erythropoietin expression attenuates neuronal damage under hypothermic conditions J Neuroinflammation, 2020-05-02;17(1):141. 2020-05-02 [PMID: 32359362] (Western Blot, Rat)
• A Morin, J Goncalves, S Moog, LJ Castro-Veg, S Job, A Buffet, MJ Fontenille, J Woszczyk, AP Gimenez-Ro, E Letouzé, J Favier TET-Mediated Hypermethylation Primes SDH-Deficient Cells for HIF2&amp;alpha-Driven Mesenchymal Transition Cell Rep, 2020-03-31;30(13):4551-4566.e7. 2020-03-31 [PMID: 32234487] (ICC, Mouse)
• Christina Wohlrab, Caroline Kuiper, Margreet CM Vissers, Elisabeth Phillips, Bridget A Robinson, Gabi U Dachs <p>Ascorbate modulates the hypoxic pathway by increasing intracellular activity of the HIF hydroxylases in renal cell carcinoma cells</p> Hypoxia (Auckl) 2019-05-15 [PMID: 31192266]
• P Zhang, L Cao, R Zhou, X Yang, M Wu The lncRNA Neat1 promotes activation of inflammasomes in macrophages Nat Commun, 2019-04-02;10(1):1495. 2019-04-02 [PMID: 30940803] (Western Blot, Mouse)
• DT Dao, L Anez-Busti, SS Jabbouri, A Pan, H Kishikawa, PD Mitchell, GL Fell, MA Baker, RS Watnick, H Chen, MS Rogers, DR Bielenberg, M Puder A paradoxical method to enhance compensatory lung growth: Utilizing a VEGF inhibitor PLoS ONE, 2018-12-19;13(12):e0208579. 2018-12-19 [PMID: 30566445] (IHC-P, Mouse)
• Christina Wohlrab, Margreet C. M. Vissers, Elisabeth Phillips, Helen Morrin, Bridget A. Robinson, Gabi U. Dachs The Association Between Ascorbate and the Hypoxia-Inducible Factors in Human Renal Cell Carcinoma Requires a Functional Von Hippel-Lindau Protein Frontiers in Oncology 2018-11-30 [PMID: 30555801]
• Christopher J. Greene, Nitika J. Sharma, Peter N. Fiorica, Emily Forrester, Gary J. Smith, Kenneth W. Gross et al. Suppressive effects of iron chelation in clear cell renal cell carcinoma and their dependency on VHL inactivation Free Radical Biology and Medicine 2019-03-01 [PMID: 30553971] (Western Blot, Human)
• Cindy V Leiton, Elyssa Chen, Alissa Cutrone, Kristy Conn, Kennelia Mellanson, Dania M Malik et al. Astrocyte HIF-2&alpha; supports learning in a passive avoidance paradigm under hypoxic stress Hypoxia (Auckl) 2018-12-07 [PMID: 30519596]
• Schwartz AJ, Das Nk, Ramakrishnan Sk et al. Hepatic hepcidin/intestinal HIF-2a axis maintains iron absorption during iron deficiency and overload. J. Clin. Invest. 10/23/2018 [PMID: 30352047]
• Yi-Feng Gu, Shannon Cohn, Alana Christie, Tiffani McKenzie, Nicholas Wolff, Quyen N. Do et al. Modeling Renal Cell Carcinoma in Mice: Bap1 and Pbrm1 Inactivation Drive Tumor Grade Cancer Discovery 2017-08-01 [PMID: 28473526]
• Guentsch A, Beneke A, Swain L et al. PHD2 Is a Regulator for Glycolytic Reprogramming in Macrophages. Mol. Cell. Biol. 1/1/2017 [PMID: 27795296]
• Semba H, Takeda N, Isagawa T et al. HIF-1alpha-PDk1 axis-induced active glycolysis plays an essential role in macrophage migratory capacity. Nat Commun. 5/18/2016 [PMID: 27189088]
• H Kobayashi, Q Liu, TC Binns, AA Urrutia, O Davidoff, PP Kapitsinou, AS Pfaff, H Olauson, A Wernerson, AB Fogo, GH Fong, KW Gross, VH Haase Distinct subpopulations of FOXD1 stroma-derived cells regulate renal erythropoietin J Clin Invest, 2016-04-18;0(0):. 2016-04-18 [PMID: 27088801] (Western Blot, Mouse)
• Milora K, Fu H, Dubaz O, Jensen L Unprocessed Interleukin-36alpha Regulates Psoriasis-Like Skin Inflammation in Cooperation With Interleukin-1. J Invest Dermatol, 2015-07-23;135(12):2992-3000. 2015-07-23 [PMID: 26203636] (ELISA Development, Mouse)
• Mangiavini L, Merceron C, Araldi E, Khatri R, Gerard-O'Riley R, Wilson T, Rankin E, Giaccia A, Schipani E Loss of VHL in mesenchymal progenitors of the limb bud alters multiple steps of endochondral bone development. Dev Biol, 2014-06-24;393(1):124-36. 2014-06-24 [PMID: 24972088] (Western Blot, Mouse)
• Swain L, Wottawa M, Hillemann A, Beneke A, Odagiri H, Terada K, Endo M, Oike Y, Farhat K, Katschinski D Prolyl-4-hydroxylase domain 3 (PHD3) is a critical terminator for cell survival of macrophages under stress conditions. J Leukoc Biol, 2014-03-13;96(3):365-75. 2014-03-13 [PMID: 24626957] (Western Blot, Mouse)
• Weidemann A, Breyer J, Rehm M, Eckardt K, Daniel C, Cicha I, Giehl K, Goppelt-Struebe M HIF-1alpha activation results in actin cytoskeleton reorganization and modulation of Rac-1 signaling in endothelial cells. Cell Commun Signal, 2013-10-21;11(0):80. 2013-10-21 [PMID: 24144209] (Western Blot, Mouse)
• Dehne N, Tausendschon M, Essler S, Geis T, Schmid T, Brune B IL-4 reduces the proangiogenic capacity of macrophages by down-regulating HIF-1alpha translation. J Leukoc Biol, 2013-09-04;95(1):129-37. 2013-09-04 [PMID: 24006507] (Western Blot, Human)
• Wottawa M, Leisering P, Ahlen M, Schnelle M, Vogel S, Malz C, Bordoli M, Camenisch G, Hesse A, Napp J, Alves F, Kristiansen G, Farhat K, Katschinski D Knockdown of prolyl-4-hydroxylase domain 2 inhibits tumor growth of human breast cancer MDA-MB-231 cells by affecting TGF-beta1 processing. Int J Cancer, 2012-12-27;132(12):2787-98. 2012-12-27 [PMID: 23225569] (Western Blot, Human)
• Aro E, Khatri R Hypoxia-inducible Factor-1 (HIF-1) but Not HIF-2 Is Essential for Hypoxic Induction of Collagen Prolyl 4-Hydroxylases in Primary Newborn Mouse Epiphyseal Growth Plate Chondrocytes. J. Biol. Chem., 2012-08-28;287(44):37134-44. 2012-08-28 [PMID: 22930750] (Western Blot, Mouse)
• Liang D, Ma Y, Liu J, Trope CG, Holm R, Nesland JM, Suo Z The hypoxic microenvironment upgrades stem-like properties of ovarian cancer cells. BMC Cancer, 2012-05-29;12(0):201. 2012-05-29 [PMID: 22642602] (Western Blot, Human)
• Branco-Price C, Zhang N, Schnelle M et al. Endothelial Cell HIF-1[alpha] and HIF-2[alpha] Differentially Regulate Metastatic Success. Cancer Cell 1721(1):52-65. 1/1/2012 [PMID: 22264788]
• Norihiko Takeda, Ellen L. O'Dea, Andrew Doedens, Jung-whan Kim, Alexander Weidemann, Christian Stockmann et al. Differential activation and antagonistic function of HIF-{alpha} isoforms in macrophages are essential for NO homeostasis Genes & Development 2010-03-01 [PMID: 20194441]
• Stockmann C, Kerdiles Y, Nomaksteinsky M, Weidemann A, Takeda N, Doedens A, Torres-Collado AX, Iruela-Arispe L, Nizet V, Johnson RS Loss of myeloid cell-derived vascular endothelial growth factor accelerates fibrosis. Proc. Natl. Acad. Sci. U.S.A., 2010-02-08;107(9):4329-34. 2010-02-08 [PMID: 20142499] (Western Blot, Mouse)

Bioinformatics

• Gene Symbol: EPAS1
• Uniprot:
  • AAH57870()