Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

c-Cbl Antibody Sampler Kit #9794

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
c-Cbl (C49H8) Rabbit mAb #2179 40 µl W IP H Mk 120 Rabbit IgG
Phospho-c-Cbl (Tyr731) Antibody #3554 40 µl W H 120 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat
Phospho-c-Cbl (Tyr700) (D16D7) Rabbit mAb #8869 40 µl W IP H (M) 120 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

The Phospho-c-Cbl (Tyr700) and (Tyr731) Antibodies detect endogenous c-Cbl when phosphorylated at the specified sites. c-Cbl (C49H3) Rabbit mAb detects endogenous levels of total c-Cbl protein.

Western Blotting

Western Blotting

Western blot analysis of cell extracts from various cell types using c-Cbl (C49H8) Rabbit mAb #2179.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293-T cells transfected with c-Cbl and Fyn (lanes 3 and 6) or without Fyn (lanes 2 and 5), untreated or calf intestinal phosphatase (CIP)-treated, using Phospho-c-Cbl (Tyr731) Antibody #3554. (Lysates provided by Dr. A.L. Reddi, laboratory of Dr. Hamid Band, Harvard University, Massachusetts.)

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated (-) or treated with pervanadate (1 mM; +), using Phospho-c-Cbl (Tyr700) (D16D7) Rabbit mAb #8869 (upper) or c-Cbl (D4E10) Rabbit mAb #8447 (lower).


Description

The c-Cbl Antibody Sampler Kit provides an economical means of evaluating levels of c-Cbl protein phosphorylated at the specified sites, as well as total c-Cbl levels. The kit contains enough reagents to perform four western blot experiments per primary antibody.

Source / Purification

The phospho-specific polyclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr731 of human c-Cbl. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. The monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to the sequence of human c-Cbl and residues surrounding Tyr700 of human c-Cbl.

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).

  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.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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