Key features and details
- Rabbit polyclonal to HIF-2-alpha
- Suitable for: WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Product nameAnti-HIF-2-alpha antibody
See all HIF-2-alpha primary antibodies
DescriptionRabbit polyclonal to HIF-2-alpha
Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Chimpanzee, Macaque monkey, Chinese hamster, Orangutan
Synthetic peptide corresponding to Human HIF-2-alpha aa 350-450 conjugated to keyhole limpet haemocyanin.
- WB: Human placenta and heart tissue lysates and A549, HeLa, NIH3T3 and PC12 whole cell lysates. IHC: Human colon tissue. ICC/IF: HeLa cells (untreated and treated with Deferoxamine).
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Concentration information loading...
PurityImmunogen affinity purified
- Pathways and Processes
- Metabolic signaling pathways
- Nucleotide metabolism
- Molecular processes
- Mitochondrial transcription
Our Abpromise guarantee covers the use of ab109616 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 96 kDa).|
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.|
|ICC/IF||Use a concentration of 10 µg/ml.|
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
- Basic helix loop helix PAS protein MOP2 antibody
- Basic-helix-loop-helix-PAS protein MOP2 antibody
- bHLHe73 antibody
ab109616 staining Hif-2-alpha in HeLa cells. The cells were incubated with 1mM Deferoxamine for 24 hours (Treated) or solvent-only for control purposes (Solvent only). Cells were fixed with 4% formaldehyde (10min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab109616 at 10µg/ml and ab195889 at 1µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.
IHC image of HIF-2alpha staining in a section of formalin-fixed paraffin-embedded normal human colon*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 for 20mins. The section was then incubated with ab109616 at 1μg/ml, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
All lanes : Anti-HIF-2-alpha antibody (ab109616) at 1 µg/ml (Milk blocking - 3%)
Lane 1 : Human placenta tissue lysate - total protein (ab29745)
Lane 2 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate
Lane 3 : Human heart tissue lysate - total protein (ab29431)
Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 6 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 96 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?
Additional bands at: 100 kDa, 38 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 16 minutes
The predicted molecular weight of HIF2 alpha is 96 kDa (SwissProt), however we expect to observe a banding pattern around 120 kDa. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
ab109616 has been referenced in 4 publications.
- Rosenblum JS et al. Neuraxial dysraphism in EPAS1-associated syndrome due to improper mesenchymal transition. Neurol Genet 6:e414 (2020). PubMed: 32337341
- Mira E et al. SOD3 improves the tumor response to chemotherapy by stabilizing endothelial HIF-2a. Nat Commun 9:575 (2018). WB . PubMed: 29422508
- He S et al. DKK4 enhances resistance to chemotherapeutics 5-Fu and YN968D1 in colorectal cancer cells. Oncol Lett 13:587-592 (2017). WB . PubMed: 28356933
- D'Hulst G et al. Fifteen days of 3,200 m simulated hypoxia marginally regulates markers for protein synthesis and degradation in human skeletal muscle. Hypoxia (Auckl) 4:1-14 (2016). PubMed: 27800505