The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use 2µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Use at an assay dependent concentration.
Use at an assay dependent concentration.
1/1000 - 1/3000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
1/100 - 1/500. Detects a band of approximately 118 kDa (predicted molecular weight: 100 kDa).
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.
Expressed in most tissues, with highest levels in placenta, lung and heart. Selectively expressed in endothelial cells.
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.
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.
Class E basic helix-loop-helix protein 73 antibody
Endothelial PAS domain containing protein 1 antibody
Endothelial pas domain protein 1 antibody
Endothelial PAS domain-containing protein 1 antibody
EPAS 1 antibody
HIF 1 alpha like factor antibody
HIF 2 alpha antibody
HIF-1-alpha-like factor 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
PAS domain-containing protein 2 antibody
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF-2-alpha antibody [ep190b] (ab8365)Image from Xiang L et al., Diagn Pathol. 2012 Mar 27;7:32. Fig 5.; doi:10.1186/1746-1596-7-32; 27 March 2012, Diagnostic Pathology 2012, 7:32
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.
Western blot - Anti-HIF-2-alpha antibody [ep190b] (ab8365)
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.
Polívka J et al. IDH1 mutation is associated with lower expression of VEGF but not microvessel formation in glioblastoma multiforme. Oncotarget9:16462-16476 (2018).
Read more (PubMed: 29662659) »
Zhang T et al. Angiopoietin-like 4 promotes osteosarcoma cell proliferation and migration and stimulates osteoclastogenesis. BMC Cancer18:536 (2018).
Read more (PubMed: 29739381) »