Product Pathways - Tyrosine Kinase / Adaptors
Phospho-CrkII (Tyr221) Antibody #3491
PhosphoSitePlus® protein, site, and accession data: Crk
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W | H Hm | Endogenous | 42 | Rabbit |
Applications Key:
W=Western Blotting
Reactivity Key:
H=Human
Hm=Hamster
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
- 3491:
- Western Blotting
Specificity / Sensitivity
Phospho-CrkII (Tyr221) Antibody detects endogenous levels of CrkII only when phosphorylated at tyrosine 221. The antibody cross-reacts with Tyr207-phosphorylated CrkL but does not cross-react with other tyrosine-phosphorylated proteins.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr221 of human CrkII. Antibodies are purified by protein A and peptide affinity chromatography.
Background
CrkII, a cellular homologue of v-Crk, belongs to a family of adaptor proteins with an SH2-SH3-SH3 domain structure that transmits signals from tyrosine kinases (1). The primary function of Crk is to recruit cytoplasmic proteins in the vicinity of tyrosine kinases through SH2-phospho-tyrosine interaction. Thus, the output from Crk depends on the SH3-binding proteins, which include the C3G and Sos guanine nucleotide exchange proteins, Abl tyrosine kinase, DOCK180 and some STE20-related kinases. The variety of Crk-binding proteins indicates the pleiotropic function of Crk (2). The two CrkII SH3 domains are separated by a 54 amino acid linker region, which is highly conserved in Xenopus, chicken and mammalian CrkII proteins (3). Tyrosine 221 in this region is phosphorylated by the Abl tyrosine kinase (4), IGF-I receptor (5) and EGF receptor (6). Once Tyr221 is phosphorylated, CrkII undergoes a change in intramolecular folding and SH2-pTyr interaction, which causes rapid dissociation of CrkII from the tyrosine kinase complex (3).
- Zvara, A. et al. (2001) Oncogene 20, 951-961.
- Kiyokawa, E. et al. (1997) Crit. Rev. Oncog. 8, 329-342.
- Rosen, S. K. et al. (1995) Nature 374, 477-479.
- Amoui, M. and Miller, W.T. (2000) Cell. Signal. 12, 637-643.
- Koval, A. P. et al. (1998) Biochem. J. 330, 923-932.
- Hashimoto, Y. et al. (1998) J. Biol. Chem. 273, 17186-17191.
Application References
- Burton, E. A. et al. (2003) Abl tyrosine kinases are required for infection by Shigella flexneri. EMBO J. 22, 5471-5479. Applications: Western Blotting
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For Research Use Only. Not For Use In Diagnostic Procedures.