Overview

  • Product nameAnti-HIF-2-alpha antibody [ep190b]
    See all HIF-2-alpha primary antibodies
  • Description
    Mouse monoclonal [ep190b] to HIF-2-alpha
  • SpecificityThis antibody is specific for HIF-2-alpha and does not cross-react with HIF-1-alpha.
  • Tested applicationsSuitable for: Flow Cyt, ELISA, IHC-Fr, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Rat, Human
  • Immunogen

    Fusion protein (proprietary-tag) corresponding to Human HIF-2-alpha aa 535-631.

  • Positive control
    • Hypoxic A549 lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab8365 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt Use 2µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ELISA Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
IHC-P 1/1000 - 1/3000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/100 - 1/500. Detects a band of approximately 118 kDa (predicted molecular weight: 100 kDa).

Target

  • FunctionTranscription factor involved in the induction of oxygen regulated genes. Binds to core DNA sequence 5'-[AG]CGTG-3' within the hypoxia response element (HRE) of target gene promoters. Regulates the vascular endothelial growth factor (VEGF) expression and seems to be implicated in the development of blood vessels and the tubular system of lung. May also play a role in the formation of the endothelium that gives rise to the blood brain barrier. Potent activator of the Tie-2 tyrosine kinase expression. Activation seems to require recruitment of transcriptional coactivators such as CREBPB and probably EP300. Interaction with redox regulatory protein APEX seems to activate CTAD.
  • Tissue specificityExpressed in most tissues, with highest levels in placenta, lung and heart. Selectively expressed in endothelial cells.
  • Involvement in diseaseDefects in EPAS1 are the cause of erythrocytosis familial type 4 (ECYT4) [MIM:611783]. ECYT4 is an autosomal dominant disorder characterized by increased serum red blood cell mass, elevated hemoglobin concentration and hematocrit, and normal platelet and leukocyte counts.
  • Sequence similaritiesContains 1 basic helix-loop-helix (bHLH) domain.
    Contains 1 PAC (PAS-associated C-terminal) domain.
    Contains 2 PAS (PER-ARNT-SIM) domains.
  • Post-translational
    modifications
    In normoxia, is probably hydroxylated on Pro-405 and Pro-531 by EGLN1/PHD1, EGLN2/PHD2 and/or EGLN3/PHD3. The hydroxylated prolines promote interaction with VHL, initiating rapid ubiquitination and subsequent proteasomal degradation. Under hypoxia, proline hydroxylation is impaired and ubiquitination is attenuated, resulting in stabilization.
    In normoxia, is hydroxylated on Asn-847 by HIF1AN thus probably abrogating interaction with CREBBP and EP300 and preventing transcriptional activation.
    Phosphorylated on multiple sites in the CTAD.
    The iron and 2-oxoglutarate dependent 3-hydroxylation of asparagine is (S) stereospecific within HIF CTAD domains.
  • Cellular localizationNucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Basic helix loop helix PAS protein MOP2 antibody
    • Basic-helix-loop-helix-PAS protein MOP2 antibody
    • bHLHe73 antibody
    • Class E basic helix-loop-helix protein 73 antibody
    • ECYT4 antibody
    • Endothelial PAS domain containing protein 1 antibody
    • Endothelial pas domain protein 1 antibody
    • Endothelial PAS domain-containing protein 1 antibody
    • EPAS 1 antibody
    • EPAS-1 antibody
    • EPAS1 antibody
    • EPAS1_HUMAN antibody
    • HIF 1 alpha like factor antibody
    • HIF 2 alpha antibody
    • HIF-1-alpha-like factor antibody
    • HIF-2-alpha antibody
    • HIF2-alpha antibody
    • HIF2A antibody
    • HLF antibody
    • Hypoxia inducible factor 2 alpha antibody
    • Hypoxia inducible factor 2 alpha subunit antibody
    • Hypoxia-inducible factor 2-alpha antibody
    • Member of PAS protein 2 antibody
    • Member of pas superfamily 2 antibody
    • MOP 2 antibody
    • MOP2 antibody
    • PAS domain-containing protein 2 antibody
    • PASD2 antibody
    see all

Anti-HIF-2-alpha antibody [ep190b] images

  • Immunohistochemical analysis of Human breast cancer tissue, staining HIF-2-alpha with ab8365.

    Antigen retrieval was performed using EDTA buffer in a pressure cooker, before being blocked with normal serum. Samples were incubated with primary antibody (1/2000) overnight at 4°C. Staining was detected using DAB.

  • ab8365 at 1/300 dilution, staining HIF-2-alpha in cardiomyocytes of Human heart tissue section by Immunohistochemistry (Formalin/PFA fixed paraffin-embedded sections).



  • Predicted band size : 100 kDa

    HIF-2-alpha detected in hypoxic Human lysate using ab8365. Lane 1: normoxic A549 lysate control, lane 2: hypoxic A549 lysate.

  • Overlay histogram showing HepG2 cells stained with ab8365 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab8365, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References for Anti-HIF-2-alpha antibody [ep190b] (ab8365)

This product has been referenced in:
  • Song W  et al. Infiltrating neutrophils promote renal cell carcinoma progression via VEGFa/HIF2a and estrogen receptor ß signals. Oncotarget 6:19290-304 (2015). WB, IHC-P . Read more (PubMed: 26079540) »
  • Trávnícek I  et al. Tissue Biomarkers in Predicting Response to Sunitinib Treatment of Metastatic Renal Cell Carcinoma. Anticancer Res 35:5661-6 (2015). Human . Read more (PubMed: 26408740) »

See all 32 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Peritonei tumor)
Specification Peritonei tumor
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative Paraformaldehyde
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Submitted Jan 19 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Tumor tisse)
Permeabilization No
Specification Tumor tisse
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative Paraformaldehyde
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Submitted Sep 29 2015

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (4-12% acrylamide)
Sample Human Cell lysate - whole cell (Renal cancer cell line)
Specification Renal cancer cell line
Blocking step Li-cor Blocking Buffer PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Jul 28 2014


I was wondering, did you try any other antigen retrieval step, such as Tris EDTA (pH 8)? This has been successfully used by another researcher with human breast cancer tissue. I would suggest trying this.

I am not sure how long you incuba...

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Thank you for contacting us.

We guarantee ab8365 to work in Flow Cyt, ELISA, IHC-Fr, IHC-P, WB with rat and human samples in accordance with our Abpromise. If you have difficulty in these applications and species, let us know and we will ref...

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Thank you for your patience whilst I have been trying to contact our supplier to get the protocol they used to test ab8365 in IHC-Fr. Unfortunately they are unable to provide that information, therefore I think the best course of action is to either re...

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Thank you for your enquiry and your interest in our products.

I can confirm that this antibody (ab8365: Anti-HIF2 alpha antibody) has been tested for IHC-Fr, IHC-P, WB, please take a look at the datasheet under tested application and the imag...

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Thank you very much for your answer. I am sorry, I think I am also a little bit confused with the HIF proteins. There are indeed different studies - on particular study showing that in MEFs (i have not found yet a study on HEK cells) the localisation o...

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Thank you for taking time to contact us. I am sorry to hear that this antibody is not providing satisfactory results. Having reviewed this case, I would like to offer some suggestions to help optimize the results with this antibody. Indeed, HIF 2 alpha...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK 293)
Loading amount 20 µg
Specification HEK 293
Gel Running Conditions Non-reduced Denaturing (7.5)
Blocking step Milk as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 21°C
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Submitted Aug 23 2011

1-10 of 22 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"