Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-EphA2 (Tyr594) Antibody #3970

Applications Reactivity Sensitivity MW (kDa) Source
W H Transfected Only 135 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-EphA2 (Tyr594) Antibody detects transfected levels of EphA2 proteins only when phosphorylated on Tyr594. This antibody does not cross-reacted with other activated protein tyrosine kinases.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr594 of human EphA2. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of cell extracts from 293-T cells, transfected with human EphA2 expression construct, untreated or stimulated with B61-Fc ligand, using Phospho-EphA2 (Tyr594) Antibody (upper) or EphA2 Antibody #3974 (lower).

Background

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 Tyr594 was identified in several tumor cell lines (8,9). It was demonstrated that phosphorylated Tyr588 and Tyr594 of EphA2 provide binding sites for guanine nucleotide exchange factors Vav2 and Vav3, which may be involved in regulation of cell migration (10).

  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. Guo, A. et al. (2008) Proc Natl Acad Sci USA 105, 692-697.
  9. Rikova, K. et al. (2007) Cell 131, 1190-1203.
  10. Fang, W.B. et al. (2008) J. Biol. Chem. 283, 16017-16026.

Application References

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Companion Products

Components of this product are covered by US Patent No. 7,807,789 owned by Cell Signaling Technology, Inc.

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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