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, IP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

120

Source/Isotype:

Rabbit IgG

UniProt ID:

#P22681

Entrez-Gene Id:

867

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Phospho-c-Cbl (Tyr700) (D16D7) Rabbit mAb recognizes endogenous levels of c-Cbl protein only when phosphorylated at Tyr700.

Species Reactivity:

Human

Species predicted to react based on 100% sequence homology

Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr700 of human c-Cbl protein.

Background

The c-Cbl proto-oncogene is a ubiquitously expressed cytoplasmic adaptor protein that is especially predominant in hematopoietic cells (1,2). c-Cbl is rapidly tyrosine-phosphorylated in response to stimulation of a variety of cell-surface receptors and becomes associated with a number of intracellular signaling molecules such as protein tyrosine kinases, phosphatidylinositol-3 kinase, Crk, and 14-3-3 proteins (3,4). c-Cbl possesses a highly conserved amino-terminal phosphotyrosine binding domain (TKB) and a C3HC4 RING finger motif. The TKB recognizes phosphorylated tyrosines on activated receptor tyrosine kinases (RTKs) as well as other nonreceptor tyrosine kinases. The RING finger motif recruits ubiquitin-conjugating enzymes. These two domains are primarily responsible for the ubiquitin ligase activity of c-Cbl and downregulation of RTKs (3). Research studies have indicated that in human cancer tissues, c-Cbl is frequently tyrosine-phosphorylated in a tumor-specific manner (5). Phosphorylation of Tyr731 of c-Cbl provides a docking site for downstream signaling components such as p85 and Fyn (6).
It has been demonstrated that c-Cbl is phosphorylated at Tyr700 by Fyn, Yes, and Syk (4) and that Vav, a hematopoietic-restricted Rac guanine nucleotide exchange factor, undergoes c-Cbl-dependent ubiquitination upon recruitment to phospho-Tyr700 (5).

  1. Blake, T.J. et al. (1991) Oncogene 6, 653-657.
  2. Thien, C.B. and Langdon, W.Y. (1998) Immunol. Cell Biol. 76, 473-482.
  3. Christine, B.F. et al. (2001) Nat. Rev. Mol. Cell Biol. 2, 294-307.
  4. Feshchenko, E.A. et al. (1998) J. Biol. Chem. 273, 8323-8331.
  5. Kamei, T. et al. (2000) Int. J. Oncol. 17, 335-339.
  6. Hunter, C. et al. (1999) J. Biol. Chem. 274, 2097-2106.
  7. Miura-Shimura, Y. et al. (2003) J Biol Chem 278, 38495-504.

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 IP: Immunoprecipitation

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.

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