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EPAS1 antibody (Endothelial PAS Domain Protein 1)

Details for Product anti-EPAS1 Antibody No. ABIN151063, Supplier: Log in to see
Antigen
  • EPAS1
  • ECYT4
  • HIF2A
  • HLF
  • MOP2
  • PASD2
  • bHLHe73
  • HIF-2alpha
  • HRF
  • Hif2a
  • bhlhe73
  • ecyt4
  • hif2
  • hif2a
  • hif2alpha
  • hlf
  • mop2
  • pasd2
  • EPAS
  • EPAS-1
  • endothelial PAS domain protein 1
  • EPAS1
  • epas1
  • Epas1
Reactivity
Hamster, Human, Mouse (Murine), Rat (Rattus)
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Host
Mouse
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1
Clonality (Clone)
Monoclonal ()
Conjugate
This EPAS1 antibody is un-conjugated
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Application
Chromatin Immunoprecipitation (ChIP), ELISA, Flow Cytometry (FACS), Gel Shift (GS), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP), Simple Western (SimWes), Western Blotting (WB)
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Options
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Immunogen Human HIF-2 alpha, corresponding to amino acids 535-631. [UniProt# Q99814]
Clone Ep190b
Isotype IgG1
Specificity This is specific for HIF-2 alpha and does not cross-react with HIF-1 alpha.
Cross-Reactivity (Details) Ability to use in mouse is mixed with some positive,some negative results.
Purification Protein G purified
Alternative Name HIF-2 alpha/epas1 (EPAS1 Antibody Abstract)
Background Gene Symbol: EPAS1
Molecular Weight Theoretical MW: 118 kDa
Gene ID 2034
UniProt Q99814
Application Notes Western Blot 1-2 μg/mL, Simple Western 1:100, Chromatin Immunoprecipitation, Flow Cytometry 1:400, ELISA 1:100-1:2000, Immunohistochemistry 1:150-1:300, Immunocytochemistry/Immunofluorescence, Immunoprecipitation, Immunohistochemistry-Paraffin 1:150-1:300, Gel Supershift AssayThis 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) In Simple Western only 10 - 15 μL 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.
Comment

The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.

Protocol Protocol specific for HIF-2 alpha Antibody Procedure Guide for NB 100-132Monoclonal Anti-HIF-2 alpha Western Blot Procedure
1. Resolve nuclear cell extracts (50-100 µg/lane) on a 6 % SDS-polyacrylamide gel, under reducing conditions.
. Transfer to a nitrocellulose membrane, overnight, or to a *PVDF membrane [*in 20 mM Tris/100 mM glycine/10 % (v/v) methanol/0.05 % SDS].
. Block the membrane in TBS containing 5 % non-fat dry milk and 0.1 % Tween-20.
. Rinse the membrane in TBST, twice.
. Incubate the membrane in anti-HIF-2 alpha , diluted 1:500 in TBS+1 % BSA, overnight at 4C.
. Wash membrane with TBST for 35 minutes at RT (1 X 15 minutes, 2 X 10 minutes).
. Incubate the membrane with diluted HRP conjugated goat anti-mouse antibody.
. Wash membrane with TBST for 35 minutes at RT (1 X 15 minutes, 2 X 10 minutes).
. Use Amersham ECL Kit, as directed, to detect image.Immunohistochemistry Procedure for Paraffin Sections
. Prior to performing the IPOX experiment, dewax the paraffin sections by baking them at 60C for 30 minutes and then putting them through citroclear.
. Hydrate the sections through the following series:A. 3 X 5 minutes xylenesB. 3 X 5 minutes 100 % EtohC. 2 minutes 95 % EtohD. 2 minutes 70 % EtohE. 1 minute 50 % EtohF. 1 minute ddH2OG. 1 minute TBS
. Block endogenous peroxidase with 0.5 % hydrogen peroxide in water, for 30 minutes.
. Antigen unmasking is performed by incubating at 60C for 16 hours, in 50mmol/L Tris and 0.2 mmol/L EDTA (pH 9.0), using a covered water bath.
. Rinse slides with PBS and then incubate with PBS containing 0.2 % Triton X-100 for 10 minutes.
. Rinse slides with PBS.
. Incubate sections with 1:1,000-1:3,000 dilution of anti-HIF-2 alpha for 90 minutes at RT.
. Incubate sections in secondary HRP-conjugated goat anti-mouse serum for 30 minutes at RT.
. Incubate sections in tertiary HRP-conjugated rabbit anti-goat serum for 30 minutes at RT.
. Develop the peroxidase reaction using diaminobenzidine.
. Wash slide and mount in aqueous mountant.Substitution of the primary antibody with PBS can be used as a negative control.
. Sub-confluent cells are grown on chamber slides and incubated for 16 hours either in air or under 0.1 % hypoxia.
. Wash cells in ice-cold PBS.
. Fix cells in formaldehyde (3.7 % in PBS) for 10 minutes at room temperature (RT).
. Wash cells twice, in PBS, and permeabilize by incubating in 0.2 % Triton X-100 in PBS for 10 minutes at RT.
. Incubate the slides with 1:1,000-1:3,000 dilution of anti-HIF-2 alpha for 1 hour at RT.
. Wash in PBS for 5 minutes.
. Incubate with HRP-conjugated goat anti-mouse for 30 minutes at RT.
. Detect binding using 3Prime-diaminobenzidine.
. Counterstain with hematoxylin.IHC-FFPE sectionsI. Deparaffinization: A. Treat slides with Xylene: 3 changes for 5 minutes each. Drain slides for 10 seconds between changes. B. Treat slides with 100 % Reagent Alcohol: 3 changes for 5 minutes each. Drain slides for 10 seconds between changes.II. Quench Endogenous Peroxidase: A. Place slides in peroxidase quenching solution: 15-30 minutes. To Prepare 200 mL of Quenching Solution: -Add 3 mL of 30 % Hydrogen Peroxide to 200 mL of Methanol.
Restrictions For Research Use only
Format Liquid
Concentration 1.0 mg/mL
Buffer PBS with 1 % BSA
Buffer contains: 0.05 % Sodium Azide
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Avoid freeze-thaw cycles
Storage -80 °C,-20 °C
Storage Comment Aliquot and store at -20°C or -80°C. Avoid freeze-thaw cycles.
Supplier Images
Western Blotting (WB) image for anti-EPAS1 antibody (Endothelial PAS Domain Protein 1) (ABIN151063) HIF-2 alpha detected in hypoxic human lysate using ABIN151063. Lane 1: normoxic A549 ...
Immunohistochemistry (IHC) image for anti-EPAS1 antibody (Endothelial PAS Domain Protein 1) (ABIN151063) Staining of heart, cardiac myocytes using ABIN151063.
Western Blotting (WB) image for anti-EPAS1 antibody (Endothelial PAS Domain Protein 1) (ABIN151063) Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [ABIN151064] - HRP conjugated HIF-2...
Simple Western (SimWes) image for anti-EPAS1 antibody (Endothelial PAS Domain Protein 1) (ABIN151063) Simple Western: HIF-2 alpha/EPAS1 Antibody (ep190b) [ABIN151064] - Simple Western lan...
Flow Cytometry (FACS) image for anti-EPAS1 antibody (Endothelial PAS Domain Protein 1) (ABIN151063) Flow Cytometry: HIF-2 alpha/EPAS1 Antibody (ep190b) [ABIN151064] - HIF-2 alpha antibo...
Western Blotting (WB) image for anti-EPAS1 antibody (Endothelial PAS Domain Protein 1) (ABIN151063) Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [ABIN151064] - Western blot analysi...
Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)) image for anti-EPAS1 antibody (Endothelial PAS Domain Protein 1) (ABIN151063) Immunohistochemistry-Paraffin: HIF-2 alpha/EPAS1 Antibody (ep190b) [ABIN151064] - Ana...
Western Blotting (WB) image for anti-EPAS1 antibody (Endothelial PAS Domain Protein 1) (ABIN151063) Western Blot: HIF-2 alpha/EPAS1 Antibody (ep190b) [ABIN151064] - C57B6 5 week old Mou...
Immunofluorescence (IF) image for anti-EPAS1 antibody (Endothelial PAS Domain Protein 1) (ABIN151063) Immunofluorescence: HIF-2 alpha/EPAS1 Antibody (ep190b) [ABIN151064] - Analysis of HI...
Product cited in: Seidl, Martinez-Sanchez, Murphy: "Derepression of MicroRNA-138 Contributes to Loss of the Human Articular Chondrocyte Phenotype." in: Arthritis & rheumatology (Hoboken, N.J.), Vol. 68, Issue 2, pp. 398-409, 2016 (PubMed). (Sample species: Human). Further details: Western Blotting

Lee, Frattini, Bansal, Castano, Sherman, Hutchinson, Bruce, Califano, Liu, Cardozo, Iavarone, Lasorella: "An ID2-dependent mechanism for VHL inactivation in cancer." in: Nature, Vol. 529, Issue 7585, pp. 172-7, 2016 (PubMed). (Sample species: Human). Further details: Immunohistochemistry (Paraffin-embedded Sections)

Kim, Nam, Lee, Park, Kim, Yu, Kim, Park, Bhin, Hwang, Lee, Koh, Baek: "Methylation-dependent regulation of HIF-1α stability restricts retinal and tumour angiogenesis." in: Nature communications, Vol. 7, pp. 10347, 2016 (PubMed). Method employed by authors: Immunofluorescence (IF)

Espana-Agusti, Zou, Wong, Fu, Yang, Tuveson, Adams, Matakidou: "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." in: PLoS ONE, Vol. 11, Issue 2, pp. e0148055, 2016 (PubMed). Method employed by authors: Western Blotting (WB)

Chen, Zhou, Zhou, Tang, Ibe, Cheng, Gou, Chen, Yuan, Raj: "Loss of microRNA-17∼92 in smooth muscle cells attenuates experimental pulmonary hypertension via induction of PDZ and LIM domain 5." in: American journal of respiratory and critical care medicine, Vol. 191, Issue 6, pp. 678-92, 2015 (PubMed). (Sample species: Human). Further details: Western Blotting

Påhlman, Lund, Jögi: "Differential HIF-1α and HIF-2α Expression in Mammary Epithelial Cells during Fat Pad Invasion, Lactation, and Involution." in: PLoS ONE, Vol. 10, Issue 5, pp. e0125771, 2015 (PubMed). (Sample species: Mouse (Murine)). Further details: Immunohistochemistry (Paraffin-embedded Sections)

Wong, Richardson, Seykora, Cotsarelis, Simon: "Hypoxia-inducible factors regulate filaggrin expression and epidermal barrier function." in: The Journal of investigative dermatology, Vol. 135, Issue 2, pp. 454-61, 2015 (PubMed). (Sample species: Mouse (Murine)).

Chen, Xu, Nagati, Hogg, Das, Gerard, Garcia: "The Acetate/ACSS2 Switch Regulates HIF-2 Stress Signaling in the Tumor Cell Microenvironment." in: PLoS ONE, Vol. 10, Issue 2, pp. e0116515, 2015 (PubMed). (Sample species: Human). Further details: Western Blotting,Immunoprecipitation

Her, Nelson-Holte, Maher: "Oxygen concentration controls epigenetic effects in models of familial paraganglioma." in: PLoS ONE, Vol. 10, Issue 5, pp. e0127471, 2015 (PubMed). Further details: Western Blotting,Immunohistochemistry (Paraffin-embedded Sections)

Espana-Agusti, Tuveson, Adams, Matakidou: "A minimally invasive, lentiviral based method for the rapid and sustained genetic manipulation of renal tubules." in: Scientific reports, Vol. 5, pp. 11061, 2015 (PubMed). (Sample species: Mouse (Murine)). Further details: Immunohistochemistry

Fukui, Kageyama, Higashi, Kihara, Kizaka-Kondoh, Hiraoka, Shinojima, Suzuki, Oya: "Development of a novel interferon-α2b gene construct with a repetitive hypoxia-inducible factor binding site and its suppressive effects on human renal cell carcinoma cell lines in vitro." in: International journal of clinical oncology, Vol. 19, Issue 3, pp. 497-504, 2014 (PubMed). (Sample species: Human). Further details: Western Blotting

Thompson, Elks, Marriott, Eamsamarng, Higgins, Lewis, Williams, Parmar, Shaw, McGrath, Formenti, Van Eeden, Kinnula, Pugh, Sabroe, Dockrell, Chilvers, Robbins, Percy, Simon, Johnson, Renshaw, Whyte et al.: "Hypoxia-inducible factor 2α regulates key neutrophil functions in humans, mice, and zebrafish. ..." in: Blood, Vol. 123, Issue 3, pp. 366-76, 2014 (PubMed). Further details: Western Blotting,Immunohistochemistry

Ellinghaus, Heisler, Unterschemmann, Haerter, Beck, Greschat, Ehrmann, Summer, Flamme, Oehme, Thierauch, Michels, Hess-Stumpp, Ziegelbauer: "BAY 87-2243, a highly potent and selective inhibitor of hypoxia-induced gene activation has antitumor activities by inhibition of mitochondrial complex I." in: Cancer medicine, Vol. 2, Issue 5, pp. 611-24, 2014 (PubMed). Further details: Western Blotting

Duan, Takeda, Fong: "Hypoxia inducible factor-2α regulates the development of retinal astrocytic network by maintaining adequate supply of astrocyte progenitors." in: PLoS ONE, Vol. 9, Issue 1, pp. e84736, 2014 (PubMed). (Sample species: Mouse (Murine)). Further details: Western Blotting

Gregg, Turner, Chang, Joshi, Zhan, Chen, Maranchie: "NADPH oxidase NOX4 supports renal tumorigenesis by promoting the expression and nuclear accumulation of HIF2α." in: Cancer research, Vol. 74, Issue 13, pp. 3501-11, 2014 (PubMed). (Sample species: Human). Further details: Immunocytochemistry,Western Blotting,Immunohistochemistry (Paraffin-embedded Sections),Immunofluorescence

Toyoda, Hirayama, Sugimoto, Uchida, Ohishi, Hirayama, Komada: "Polycythemia and paraganglioma with a novel somatic HIF2A mutation in a male." in: Pediatrics, Vol. 133, Issue 6, pp. e1787-91, 2014 (PubMed). (Sample species: Human). Further details: Immunohistochemistry (Paraffin-embedded Sections)

Carrera, Senra, Acosta, Althubiti, Hammond, de Verdier, Macip: "The role of the HIF-1α transcription factor in increased cell division at physiological oxygen tensions." in: PLoS ONE, Vol. 9, Issue 5, pp. e97938, 2014 (PubMed). (Sample species: Human). Further details: Western Blotting

Evans, Wadoodi, Humphries, Lu, Grover, Saha, Smith: "Local accumulation of hypoxia-inducible factor 2 alpha during venous thrombus resolution." in: Thrombosis research, Vol. 134, Issue 3, pp. 757-60, 2014 (PubMed). (Sample species: Mouse (Murine)). Further details: Immunohistochemistry (Paraffin-embedded Sections)

Xu, Nagati, Xie, Li, Walters, Moon, Gerard, Huang, Comerford, Hammer, Horton, Chen, Garcia: "An acetate switch regulates stress erythropoiesis." in: Nature medicine, Vol. 20, Issue 9, pp. 1018-26, 2014 (PubMed). (Sample species: Human). Further details: Western Blotting,Immunoprecipitation,Chromatin Immunoprecipitation

Duan, Takeda, Fong: "Hematological, hepatic, and retinal phenotypes in mice deficient for prolyl hydroxylase domain proteins in the liver." in: The American journal of pathology, Vol. 184, Issue 4, pp. 1240-50, 2014 (PubMed). (Sample species: Mouse (Murine)). Further details: Western Blotting