Revision 1
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

SENSITIVITY:

Endogenous

MW (kDa):

145

SOURCE:

Rabbit

UniProt ID:

#P10721

Entrez-Gene Id:

3815

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-c-Kit (Tyr568/570) Antibody recognizes endogenous levels of c-Kit protein only when phosphorylated at Tyr568/570. This antibody also recognizes a hSCF-induced, non-specific band at 95 kDa from a protein of unknown identity.

Species Reactivity:

Human

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 Tyr568/570 of human c-Kit protein. Antibodies are purified by protein A and peptide affinity chromatography.

Background

c-Kit is a member of the subfamily of receptor tyrosine kinases that includes PDGF, CSF-1, and FLT3/flk-2 receptors (1,2). It plays a critical role in activation and growth in a number of cell types, including hematopoietic stem cells, mast cells, melanocytes, and germ cells (3). Upon binding with its stem cell factor (SCF) ligand, c-Kit undergoes dimerization/oligomerization and autophosphorylation. Activation of c-Kit results in the recruitment and tyrosine phosphorylation of downstream SH2-containing signaling components, including PLCγ, the p85 subunit of PI3 kinase, SHP2, and CrkL (4). Molecular lesions that impair the kinase activity of c-Kit are associated with a variety of developmental disorders (5), and mutations that constitutively activate c-Kit can lead to pathogenesis of mastocytosis and gastrointestinal stromal tumors (6). Tyr719 is located in the kinase insert region of the catalytic domain. c-Kit phosphorylated at Tyr719 binds to the p85 subunit of PI3 kinase in vitro and in vivo (7).

Tyr568 and Tyr570 are located in the juxtamembrane region of c-Kit. The phosphorylation of c-Kit at these sites is ligand induced and provides a docking site for recruitment of several adaptors/kinases, including Src, Cbl, SHP-1, APS, LNK, and SOSC6. Depending on which adaptor is bound, the outcome can lead to either cell survival and proliferation or c-Kit ubiquitination and degradation (8-13).

  1. Martin, F.H. et al. (1990) Cell 63, 203-11.
  2. Yarden, Y. et al. (1987) EMBO J 6, 3341-51.
  3. Gommerman, J.L. et al. (1997) J Biol Chem 272, 30519-25.
  4. Sattler, M. et al. (1997) J Biol Chem 272, 10248-53.
  5. Nocka, K. et al. (1990) EMBO J 9, 1805-13.
  6. Hirota, S. et al. (1998) Science 279, 577-80.
  7. Blume-Jensen, P. et al. (2000) Nat Genet 24, 157-62.
  8. Phung, B. et al. (2011) PLoS One 6, e24064.
  9. Simon, C. et al. (2008) Blood 112, 4039-47.
  10. Masson, K. et al. (2006) Biochem J 399, 59-67.
  11. Bayle, J. et al. (2004) J Biol Chem 279, 12249-59.
  12. Wollberg, P. et al. (2003) Biochem J 370, 1033-8.
  13. Kozlowski, M. et al. (1998) Mol Cell Biol 18, 2089-99.

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

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