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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.
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: Liu, Chen, Wong, Haynes, Baker, Wang: "Pseudohypoxia induced by miR-126 deactivation promotes migration and therapeutic resistance in renal cell carcinoma." in: Cancer letters, Vol. 394, pp. 65-75, 2017 (PubMed). Method employed by authors: Western Blotting (WB) (Sample species: Human).

Padró, Louie, Lananna, Krieg, Timmerman, Chan: "Genome-independent hypoxic repression of estrogen receptor alpha in breast cancer cells." in: BMC cancer, Vol. 17, Issue 1, pp. 203, 2017 (PubMed). Method employed by authors: Western Blotting (WB), Immunofluorescence (fixed cells) (IF/ICC) (Sample species: Human).

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)

Leszczynska, Dobrynin, Leslie, Ient, Boumelha, Senra, Hawkins, Maughan, Mukherjee, Hammond: "Preclinical testing of an Atr inhibitor demonstrates improved response to standard therapies for esophageal cancer." in: Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology, Vol. 121, Issue 2, pp. 232-238, 2016 (PubMed). Method employed by authors: Western Blotting (WB) (Sample species: Human).

Endo, Okami, Okuyama, Nishizawa, Imamura, Inoue: "The induction of MIG6 under hypoxic conditions is critical for dormancy in primary cultured lung cancer cells with activating EGFR mutations." in: Oncogene, Vol. 36, Issue 20, pp. 2824-2834, 2016 (PubMed). Method employed by authors: Western Blotting (WB) (Sample species: Human).

Kwak, Lee, Lee, Hong, Nam: "HIF2α/EFEMP1 cascade mediates hypoxic effects on breast cancer stem cell hierarchy." in: Oncotarget, Vol. 7, Issue 28, pp. 43518-43533, 2016 (PubMed). Method employed by authors: Western Blotting (WB) (Sample species: Mouse (Murine)).

Szendrői, Szász, Kardos, Tőkés, Idan, Szűcs, Kulka, Nyirády, Szendrői, Szállási, Győrffy, Tímár: "Opposite prognostic roles of HIF1α and HIF2α expressions in bone metastatic clear cell renal cell cancer." in: Oncotarget, Vol. 7, Issue 27, pp. 42086-42098, 2016 (PubMed). Method employed by authors: Immunohistochemistry (IHC)

Nüsken, Wohlfarth, Lippach, Rauh, Schneider, Dötsch, Nüsken: "Reduced Perinatal Leptin Availability May Contribute to Adverse Metabolic Programming in a Rat Model of Uteroplacental Insufficiency." in: Endocrinology, Vol. 157, Issue 5, pp. 1813-25, 2016 (PubMed).

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

Su, Li, Cui, Ji, Geng, Chai, Ma, Bai, Yang, Wuren, Ge, Rondina: "The Local HIF-2α/EPO Pathway in the Bone Marrow is Associated with Excessive Erythrocytosis and the Increase in Bone Marrow Microvessel Density in Chronic Mountain Sickness." in: High altitude medicine & biology, Vol. 16, Issue 4, pp. 318-30, 2015 (PubMed). (Sample species: Human).

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