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Technical Data Sheet

CK6940

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Crk Phospho-Regulation
Antibody Sampler Kit
Price

$395

Kit Components
Applications: WB = Western blot, E = ELISA, ICC = Immunocytochemistry, IP = Immunoprecipitation, IHC = Immunohistochemistry Species: H = Human, R = Rat, M = Mouse, C = Chicken, F = Fish

Kit Summary
The Crk phospho-regulation antibody sampler kit can be used to detect phosphorylation of Tyr-221 and Tyr-251 in Crk II, as well as phosphorylation of Tyr-207 and Tyr-251 in CrkL. The kit includes a rabbit polyclonal anti-CrkL and mouse monoclonal anti-Crk II antibody to detect Crk expression levels.
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Enlarge

Western blot of human K562 cells stimulated with pervanadate (1 mM) for 30 min. (lanes 1-6). The blot was treated with alkaline phosphatase to dephosphorylate Crk (lanes 2, 4 & 6), then the blot was probed with rabbit polyclonals CrkL (C-terminus) CP3081 (lanes 1 & 2), Crk (Tyr-251) phospho-specific CP3091 (lanes 3 & 4), and mouse monoclonal anti-Crk II (C-terminal region) (lanes 5 & 6).

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Enlarge

Western blot of human K562 cells stimulated with pervanadate (1 mM) for 30 min. (lanes 1-4). The blot was treated with alkaline phosphatase to dephosphorylate CrkL (lanes 2 & 4), then the blot was probed with rabbit polyclonals CrkL (C-terminus) CP3081 (lanes 1 & 2) and CrkL (Tyr-207) phospho-specific CP4671 (lanes 3 & 4).

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Background
The Crk family of adaptor proteins (Crk I, Crk II and CrkL) are Src Homology 2 (SH2) and Src Homology 3 (SH3) domain-containing proteins that form protein complexes important for transmiting signals downstream of tyrosine kinases. Both Crk II and CrkL are composed of a single SH2 domain, followed by two tandem SH3 domains. Crk II is also alternatively spliced to a minor product, Crk I, which is structurally and functionally more similar to the v-Crk oncogene. Both Crk II and CrkL are ubiquitously expressed and their SH domains are highly homologous, however both are required for mouse development and have distinct non-overlapping phenotypes in knockout mice. Phosphorylation may be important for regulating Crk activity. Crk II Tyr-221 (CrkL Tyr-207) phosphorylation is a negative regulatory site, while Crk Tyr-251 phosphorylation in the SH3 domain is a positive regulatory site. EGF stimulation induces phosphorylation of Tyr-251, which increases binding of Crk to the SH2 domain of Abl, and promotes transactivation of Abl.

Background References
Feller, S.M. (2001) Oncogene. 20(44):6348.
Feller, S.M. & Lewitzky, M. (2006) Curr. Pharm. Des. 12(5):529.
Sriram, G. et al. (2011) Oncogene 30(46):4645.
Buffer and Storage
Rabbit polyclonal and Mouse monoclonal antibodies are supplied in phosphate-buffered saline (PBS), 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store all at –20°C. Stable for 1 year.
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