Phospho-EphA2 (Tyr772) Antibod

8244:

Immunoprecipitation , Western Blotting

Phospho-EphA2 (Tyr772) Antibody recognizes endogenous levels of EphA2 protein only when phosphorylated at Tyr772. This antibody may cross-react with other overexpressed phosphotyrosine proteins.

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr772 of human EphA2. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of extracts from SNB19 cells, untreated (-) or treated (+) with EphrinA1-F_c ligand (100 ng/ml, 5 min), using Phospho-EphA2 (Tyr772) Antibody (upper) or EphA2 (D4A2) XP^® Rabbit mAb #6997 (lower).

The Eph receptors are the largest known family of receptor tyrosine kinases (RTKs). They can be divided into two groups based on sequence similarity and on their preference for a subset of ligands: EphA receptors bind to a glycosylphosphatidylinositol-anchored ephrin A ligand; EphB receptors bind to ephrin B proteins that have a transmembrane and cytoplasmic domain (1,2). Eph receptors and ligands may be involved in many diseases including cancer (3). Both ephrin A and B ligands have dual functions. As RTK ligands, the ephrins stimulate the kinase activity of the Eph receptors and activate signaling pathways in receptor-expressing cells. The ephrin extracellular domain is sufficient for this function as long as it is clustered (4). The second function of ephrins has been described as "reverse signaling", whereby the cytoplasmic domain becomes tyrosine phosphorylated, allowing interactions with other proteins that may activate signaling pathways in the ligand-expressing cells (5). Various stimuli can induce tyrosine phosphorylation of ephrin B, including binding to EphB receptors, activation of Src kinase, and stimulation by PDGF and FGF (6). Tyrosines 324/327 have been identified as major phosphorylation sites of ephrin B1 in vivo (7).

Phosphorylation of Tyr772 on EphA2 was identified at Cell Signaling Technology (CST) using PhosphoScan^® , a CST™ LC-MS/MS platform for phosphorylation site discovery (8). The phosphorylation is induced by ligand/receptor interaction (9).

1. Wilkinson, D.G. (2000) Int. Rev. Cytol. 196, 177-244.
2. Klein, R. (2001) Curr. Opin. Cell Biol. 13, 196-203.
3. Dodelet, V.C. and Pasquale, E.B. (2000) Oncogene 19, 5614-5619.
4. Holder, N. and Klein, R. (1999) Development 126, 2033-2044.
5. Brückner, K. et al. (1997) Science 275, 1640-1643.
6. Palmer, A. et al. (2002) Mol. Cell 9, 725-737.
7. Kalo, M.S. et al. (2001) J. Biol. Chem. 276, 38940-38948.
8. Rush, J. et al. (2005) Nat Biotechnol 23, 94-101.
9. Fang, W.B. et al. (2008) J Biol Chem 283, 16017-16026.

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• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 6997 EphA2 (D4A2) XP^® Rabbit mAb
• 3970 Phospho-EphA2 (Tyr594) Antibody

For Research Use Only. Not For Use In Diagnostic Procedures.