Revision 1

#3491Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB

REACTIVITY:

H Hm

SENSITIVITY:

Endogenous

MW (kDa):

42

SOURCE:

Rabbit

UniProt ID:

#P46108

Entrez-Gene Id:

1398

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

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.

Species Reactivity:

Human, Hamster

Species predicted to react based on 100% sequence homology

Mouse, Rat

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).

  1. Zvara, A. et al. (2001) Oncogene 20, 951-961.
  2. Kiyokawa, E. et al. (1997) Crit. Rev. Oncog. 8, 329-342.
  3. Rosen, M.K. et al. (1995) Nature 374, 477-9.
  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.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

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