Anti-HIF-2-alpha antibody [OTI2G5] (ab157249)
Key features and details
- Mouse monoclonal [OTI2G5] to HIF-2-alpha
- Suitable for: WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
Product nameAnti-HIF-2-alpha antibody [OTI2G5]
See all HIF-2-alpha primary antibodies
DescriptionMouse monoclonal [OTI2G5] to HIF-2-alpha
Tested applicationsSuitable for: WBmore details
Species reactivityReacts with: Human
Recombinant fragment corresponding to Human HIF-2-alpha aa 584-870.
LLDKFQQQLESKKTEPEHRPMSSIFFDAGSKASLPPCCGQASTPLSSMGG RSNTQWPPDPPLHFGPTKWAVGDQRTEFLGAAPLGPPVSPPHVSTFKTRS AKGFGARGPDVLSPAMVALSNKLKLKRQLEYEEQAFQDLSGGDPPGGSTS HLMWKRMKNLRGGSCPLMPDKPLSANVPNDKFTQNPMRGLGHPLRHLPLP QPPSAISPGENSKSRFPPQCYATQYQDYSLSSAHKVSGMASRLLGPSFES YLLPELTRYDCEVNVPVLGSSTLLQGGDLLRALDQAT
- HEK293T cells transfected with pCMV6-ENTRY HIF-2-alpha.
The clone number has been updated from 2G5 to OTI2G5, both clone numbers name the same clone.
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.30
Preservative: 0.02% Sodium azide
Constituents: 48% PBS, 50% Glycerol, 1% BSA
Concentration information loading...
PurityProtein G purified
Purification notesPurified from TCS
- Pathways and Processes
- Metabolic signaling pathways
- Nucleotide metabolism
- Molecular processes
- Mitochondrial transcription
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Our Abpromise guarantee covers the use of ab157249 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/2000. Predicted molecular weight: 96 kDa.
1/2000. Predicted molecular weight: 96 kDa.
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.
modificationsIn 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.
- Information by UniProt
- Entrez Gene: 2034 Human
- Omim: 603349 Human
- SwissProt: Q99814 Human
- Unigene: 468410 Human
- Basic helix loop helix PAS protein MOP2 antibody
- Basic-helix-loop-helix-PAS protein MOP2 antibody
- bHLHe73 antibody
All lanes : Anti-HIF-2-alpha antibody [OTI2G5] (ab157249) at 1/500 dilution
Lane 1 : Wild-type A549 Untreated (DFO Control) cell lysate
Lane 2 : Wild-type A549 Treated DFO (1 mM, 24 h) cell lysate
Lane 3 : EPAS1 knockout A549 Untreated (DFO Control) cell lysate
Lane 4 : EPAS1 knockout A549 Treated DFO (1 mM, 24 h) cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 96 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
False colour image of Western blot: Anti-HIF-2-alpha antibody [OTI2G5] staining at 1/500 dilution, shown in black; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab157249 was shown to bind specifically to HIF-2-alpha. A band was observed at 100 kDa in treated wild-type A549 cell lysates with no signal observed at this size in EPAS1 knockout cell line ab259774 (knockout cell lysate ab259779). To generate this image, wild-type and EPAS1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % BSA in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with Optiblot (ECL reagent ab133456) and imaged with 20 seconds exposure time. Secondary antibodies used were HRP conjugated Goat anti-Mouse (H+L) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
All lanes : Anti-HIF-2-alpha antibody [OTI2G5] (ab157249) at 1/2000 dilution
Lane 1 : HEK293T cells transfected with pCMV6-ENTRY control
Lane 2 : HEK293T cells transfected with pCMV6-ENTRY HIF2 alpha
Lysates/proteins at 5 µg per lane.
Predicted band size: 96 kDa
HEK293T cell lysates were generated from transient transfection of the cDNA clone (RC216194)
Datasheets and documents
ab157249 has been referenced in 6 publications.
- Hu L et al. TBK1 Is a Synthetic Lethal Target in Cancer with VHL Loss. Cancer Discov 10:460-475 (2020). PubMed: 31810986
- Hong K et al. USP37 promotes deubiquitination of HIF2a in kidney cancer. Proc Natl Acad Sci U S A N/A:N/A (2020). PubMed: 32461361
- Jiang L et al. MicroRNA-519d-3p Inhibits Proliferation and Promotes Apoptosis by Targeting HIF-2α in Cervical Cancer Under Hypoxic Conditions. Oncol Res 26:1055-1062 (2018). PubMed: 29321085
- Jiang L et al. Similarity in the functions of HIF-1a and HIF-2a proteins in cervical cancer cells. Oncol Lett 14:5643-5651 (2017). PubMed: 29098039
- Zecchini V et al. Nuclear ARRB1 induces pseudohypoxia and cellular metabolism reprogramming in prostate cancer. EMBO J 33:1365-82 (2014). WB, ICC/IF . PubMed: 24837709
- Brose SA et al. Fatty acid biosynthesis from glutamate and glutamine is specifically induced in neuronal cells under hypoxia. J Neurochem 129:400-12 (2014). PubMed: 24266789