• Product name
  • Description
    Rabbit polyclonal to HIF-2-alpha
  • Host species
  • Specificity
    ab20654 detects a HIF-2-alpha specific band at ~96 kDa in both unstimulated-PC12 cytoplasmic lysate and hypoxia-treated PC12 nuclear lysate. We do not know the identity of the additional bands observed. However, the bands at ~70 kDa (this could be HIF3 alpha) and 220 kDa have been observed with other HIF-2-alpha antibodies (Kiettzmann et al, 2001; Biochem. J. 354, pp531–537).
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide corresponding to Mouse HIF-2-alpha aa 600-700 (internal sequence) conjugated to Keyhole Limpet Haemocyanin (KLH).
    (Peptide available as ab21447, ab21447, ab21447, ab21447)

  • Positive control
    • PC12 (Rat adrenal pheochromocytoma cell line) Cytoplasmic Lysate (ab14883) PC12 (Rat adrenal pheochromocytoma cell line) Nuclear Lysate - hypoxia treated (ab14886)



Our Abpromise guarantee covers the use of ab20654 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 96 kDa (predicted molecular weight: 96 kDa).
ICC/IF Use a concentration of 1 µg/ml.
IHC-P 1/2500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.


  • Function
    Transcription 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 specificity
    Expressed in most tissues, with highest levels in placenta, lung and heart. Selectively expressed in endothelial cells.
  • Involvement in disease
    Defects 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 similarities
    Contains 1 basic helix-loop-helix (bHLH) domain.
    Contains 1 PAC (PAS-associated C-terminal) domain.
    Contains 2 PAS (PER-ARNT-SIM) domains.
  • Post-translational
    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 localization
  • 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


  • ICC/IF image of ab20654 stained rat PC12 cells. The cells were PFA fixed (10 min), permabilised in PBS-T (20 min) and incubated with the antibody (ab20654, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).


  • Image courtesy of Human Protein Atlas

    ab20654 staining HIF-2-alpha in Adrenal gland. The paraffin embedded human tissue was incubated with ab20654(1:2500 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. Ab20654 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.

    Further results for this antibody can be found at www.proteinatlas.org

  • All lanes : Anti-HIF-2-alpha antibody (ab20654) at 1 µg/ml

    Lane 1 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 2 : PC12 nuclear extract lysate (hypoxia treated) (ab14886)

    Lysates/proteins at 20 µg per lane.

    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 96 kDa
    Observed band size: 96 kDa
    Additional bands at: 240 kDa, 25 kDa, 45 kDa. We are unsure as to the identity of these extra bands.

    Exposure time: 20 minutes


This product has been referenced in:
  • Wu L  et al. Overexpression and correlation of HIF-2a, VEGFA and EphA2 in residual hepatocellular carcinoma following high-intensity focused ultrasound treatment: Implications for tumor recurrence and progression. Exp Ther Med 13:3529-3534 (2017). Read more (PubMed: 28587437) »
  • Prieto-Domínguez N  et al. Melatonin enhances sorafenib actions in human hepatocarcinoma cells by inhibiting mTORC1/p70S6K/HIF-1a and hypoxia-mediated mitophagy. Oncotarget 8:91402-91414 (2017). Read more (PubMed: 29207653) »

See all 13 Publications for this product

Customer reviews and Q&As

Thank you very much for your message Susan and for kindly sending this information to us.

I'll add it to the product details.

I appreciate your consideration. Have a great week.

Thank you for your reply.

I apologise for any confusion, I have confirmed again with our laboratories that this has not been tested in IHC-P in house. I understand this information is important for you and I am sincerely sorry we are not abl...

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Thank you for your message.

I am sorry the testing was performed by Protein Atlas in this particular case, as stated on the datasheet. We therefore regrettablyhave no further information on this occasion. I am aware however that they do the ...

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Thank you for your telephone call yesterday and for your patience. I am sorry no one had got back to you sooner.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested on th...

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Thank you for calling Abcam earlier today.

I am sorry about the issues you are having with ab20654 in IHC-P. If the protocol variations that we talked about did not prove to be effective, then please let me know. The antibody is covered under ...

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Thank you for getting back to me and sorry for the delay in my response.

Could you provide me with the document which refers to the power level 7 which you refer to? I have had some difficulty in finding a reference to this.


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Thank you for contacting us and reporting the problems you have been experiencing in using anti-HIF2 alpha antibody (ab20654) in IHC staining.

The image that you have referred to was staining of human adrenal gland which has the nucleus count...

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Thank you for contacting us. I hope you had a nice Easter weekend!

Firstly, to your question about the quantity of a drop of DABchromogen.To be honest, I am not quite sure what you are referring to with this question. Usually, a drop is the v...

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We usually recommend overnight incubation as thisallows antibodies of lower titer or affinity to be used by simply allowing more time for the antibodies to bind. Also, regardless of the antibody’s titer or affinity for its target, once the tissue...

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Thank you for contacting us.

Regarding your questions, I am happy to let you know the following:

1) Positive control for ab20654: HIF2alpha is expressed in most tissues, with highest levels in placenta, lung and heart. Also, as show...

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1-10 of 13 Abreviews or Q&A


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