Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-CrkII (Tyr221) Antibody #3491

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

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.

Western Blotting

Western Blotting

Western blot analysis of extracts from K562 cells, untreated or calf intestinal phosphatase (CIP)-treated, using Phospho-CrkII (Tyr221) Antibody.

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).

  1. Zvara, A. et al. (2001) Oncogene 20, 951-961.
  2. Kiyokawa, E. et al. (1997) Crit. Rev. Oncog. 8, 329-342.
  3. Rosen, S. K. et al. (1995) Nature 374, 477-479.
  4. Amoui, M. and Miller, W.T. (2000) Cell. Signal. 12, 637-643.
  5. Koval, A. P. et al. (1998) Biochem. J. 330, 923-932.
  6. Hashimoto, Y. et al. (1998) J. Biol. Chem. 273, 17186-17191.

Application References

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

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