Our Crk II (Tyr-221) phosphospecific primary antibody from PhosphoSolutions is rabbit polyclonal. It detects chicken, human, mouse, rat, and Xenopus Crk II (Tyr-221) and is antigen affinity purified. It is great for use in WB.
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 Crk II (lanes 2 & 4), then the blot was probed with mouse monoclonal Crk II (C-terminal region) CM3321 (lanes 1 & 2) and Crk II (Tyr-221) phospho-specific CP4701 (lanes 3 & 4).
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.
Antigen Affinity Purified
Polyclonal
IgG
WB
Rabbit
CRK
40
Phospho-Crk (Tyr-221) synthetic peptide (coupled to KLH) corresponding to amino acid residues surrounding Tyr-221 in human Crk II. This peptide sequence is well conserved in mouse and rat Crk II, and has 40% homology with the conserved site in CrkL (Tyr-207). The site is not found in Crk I.
Chicken, Frog, Human, Mouse, Rat
Storage at -20°C is recommended, as aliquots may be taken without freeze/thawing due to presence of 50% glycerol. Stable for at least 1 year at -20°C.
Liquid
PBS + 1 mg/ml BSA, 0.05% NaN3 and 50% glycerol
WB: 1:1000
Unconjugated
The antibody detects a 40 kDa* protein corresponding to the molecular mass of Crk II on SDS-PAGE immunoblots of human K562 cells stimulated with pervanadate. This reactivity is not observed after akaline phosphatase treatment.
Phosphorylated
Western blots performed on each lot.
For research use only. Not intended for therapeutic or diagnostic use. Use of all products is subject to our terms and conditions, which can be viewed on our website.
United States
After date of receipt, stable for at least 1 year at -20°C.
