Purification notesab73254 was produced from a hybridoma (mouse myeloma fused with
spleen cells from a rat immunized with recombinant mouse EphA2 extracellular domain.
The IgG fraction of culture supernatant was purified by Protein A/G affinity chromatography and lyophilized from a 0.2 µm filtered solution.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
1/100 - 1/200.
Use 1µg for 106 cells. ab18407-Rat monoclonal IgG1, is suitable for use as an isotype control with this antibody.
FunctionReceptor for members of the ephrin-A family. Binds to ephrin-A1, -A3, -A4 and -A5. Plays an important role in angiogenesis and tumor neovascularization. The recruitement of VAV2, VAV3 and PI3-kinase p85 subunit by phosphorylated EPHA2 is critical for EFNA1-induced RAC1 GTPase activation and vascular endothelial cell migration and assembly (By similarity). Induces apoptosis in a p53/TP53-independent, caspase-8-dependent manner.
Tissue specificityExpressed in brain and glioma tissue and glioma cell lines (at protein level). Expressed most highly in tissues that contain a high proportion of epithelial cells, e.g., skin, intestine, lung, and ovary.
Involvement in diseaseGenetic variations in EPHA2 are the cause of susceptibility to cataract cortical age-related type 2 (ARCC2) [MIM:613020]. A developmental punctate opacity common in the cortex and present in most lenses. The cataract is white or cerulean, increases in number with age, but rarely affects vision. Defects in EPHA2 are the cause of cataract posterior polar type 1 (CTPP1) [MIM:116600]. A subcapsular opacity, usually disk-shaped, located at the back of the lens. It can have a marked effect on visual acuity.
Sequence similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family. Ephrin receptor subfamily. Contains 2 fibronectin type-III domains. Contains 1 protein kinase domain. Contains 1 SAM (sterile alpha motif) domain.
Post-translational modificationsActivated by EFNA1 via tyrosine phosphorylation. Phosphorylated residues Tyr-588 and Tyr-594 are required for binding VAV2 and VAV3 while phosphorylated residues Tyr-735 and Tyr-930 are required for binding PI3-kinase p85 subunit. These phosphorylated residues are critical for recruitment of VAV2 and VAV3 and PI3-kinase p85 subunit which transduce downstream signaling to activate RAC1 GTPase and endothelial cell migration. They also play a critical role in transducing EPHA2 signaling in vascular endothelial cells during tumor angiogenesis.
ab73254 at 1/200 dilution, staining Eph receptor A2 in mouse kidney tissue section by Immunohistochemistry (Formalin/PFA fixed paraffin-embedded sections).
Immunoprecipitation - Eph receptor A2 antibody [RM-0051-8F21] (ab73254)This image is courtesy of an Abreview submitted by Dr. Alice Bjerregaard
ab73254 Immunoprecipitate in human A431 whole cell lysate. 500µg of cell lysate was incubated with primary antibody (undiluted in 3M ammonium sulphate, 0.1M borate buffer) and matrix (Trosylactivated Dynabeads) for 18 hours at 37°C. For western blotting an HRP-Rabbit anti-rat IgG polyclonal (1/500) was used. Lane 1: IP with 1500µg protein cell lysate. Lane 2: IP with 500µg protein cell lysate.
Overlay histogram showing A431 cells stained with ab73254 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab73254, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was goat anti-rat DyLigt 488 (IgG H+L) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgG1 [RTK2071] (ab18412, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.
References for Anti-Eph receptor A2 antibody [RM-0051-8F21] (ab73254)
has not yet been referenced specifically in any publications.
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