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.

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 rabbits with a synthetic phospho-peptide (KLH-coupled) 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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Companion Products

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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