Cell Signaling Technology

Product Pathways - Protein Stability

Cbl-b Antibody #8160

Applications Reactivity Sensitivity MW (kDa) Source
W IP H Endogenous 130 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Cbl-b Antibody recognizes endogenous levels of total Cbl-b protein. This antibody does not cross-react with other Cbl family members.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val625 of human Cbl-b protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Cbl-b Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected with a Myc/DDK-tagged cDNA expression construct encoding full-length human Cbl-b (hCbl-b-Myc/DDK, +), using Cbl-b Antibody.

Background

The Casitas B lineage lymphoma (Cbl) proteins (in mammals these are c-Cbl, Cbl-b, and Cbl-c) are a family of single subunit RING finger protein-ubiquitin E3 ligases that contain multiple protein interaction motifs (1). All Cbl proteins have a highly conserved N-terminal tyrosine kinase-binding (TKB) domain that mediates interactions between Cbl proteins and phosphorylated tyrosine residues on Cbl substrates. C-terminal to the RING finger, Cbl proteins have proline-rich domains that mediate interactions with SH3 domain-containing proteins. Phosphorylated tyrosine residues mediate interactions with SH2 domain-containing proteins such as the p85 subunit of PI3K. These protein-protein interaction motifs allow Cbl family proteins to function as adaptor proteins (2). This adaptor function contributes to the E3-dependent activities of Cbl proteins by targeting specific substrates for ubiquitination and degradation. The adaptor function also contributes to non-E3-dependent activities, such as the recruitment of proteins involved in receptor tyrosine kinase internalization, localization of Cbl proteins to specific subcellular compartments, and activation of discrete signaling pathways (1).

Cbl-b is an E3 ubiquitin ligase with a domain organization nearly identical to that of c-Cbl. The role of Cbl-b in hematopoietic cell physiology is well documented. Cbl-b expression is important for the downregulation of TCR expression during antigen recognition (2). Cbl-b also acts as a potent negative regulator of the CD28 signaling cascade to Vav and Rac1 through its ability to ubiquitinate the p85 regulatory subunit of PI3K (3,4). As a critical regulator of clonal anergy in T lymphocytes, Cbl-b mRNA and protein are upregulated in T cells following calcium mobilization and calcineurin activation (5). Cbl-b-deficient T cells are resistant to anergy induction (5). The molecular events governing this phenotype are thought to be linked to defects in the ubiquitination of PLCγ1 and PKCθ since the degradation of these signaling molecules, which occurs following restimulation of wild-type anergic T cells, fails to occur in Cbl-b-deficient T cells (5).

  1. Schmidt, M.H. and Dikic, I. (2005) Nat Rev Mol Cell Biol 6, 907-18.
  2. Naramura, M. et al. (2002) Nat Immunol 3, 1192-9.
  3. Chiang, Y.J. et al. (2000) Nature 403, 216-20.
  4. Fang, D. and Liu, Y.C. (2001) Nat Immunol 2, 870-5.
  5. Heissmeyer, V. et al. (2004) Nat Immunol 5, 255-65.

Application References

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Companion Products

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

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