Phospho-c-Cbl (Tyr774) (R4C5) rabbit mAb
Catalog #: 2361
20 μL is enough antibody for at least 20 Western blots.
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The c-Cbl (Casitas B-lineage Lymphoma) proto-oncogene is a ubiquitously expressed cytoplasmic adaptor protein that contains multiple functional domains, including an amino-terminal tyrosine kinase-binding (TKB) domain, a RING finger motif, and a proline-rich region. The TKB recognizes phosphorylated tyrosines on activated receptor tyrosine kinases (RTKs) and on other nonreceptor tyrosine kinases, while 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 (1). The proline-rich region contains 14-3-3 protein-binding and SH3 domain-binding motifs. c-Cbl is phosphorylated at Y700, Y731, and Y774 by Syk- and Src-family kinases after the stimulation of some integrins and a wide variety of receptors for immunoglobulins, antigens, hormones, growth factors, and cytokines. Phosphorylated Y774 interacts with the SH2 domain of Crk (1,2). The c-Cbl adapter protein is expressed in the cytoplasm in all tissues, with especially high levels of expression in hematopoietic cells (3,4). Through its many functional sites, c-Cbl plays key roles in the positive and negative regulation of vital cell functions, including T Cell Receptor-mediated cellular immune responses. In human cancer tissues, c-Cbl is frequently tyrosine-phosphorylated in a tumor-specific manner (5).
|Applications||Flow Cytometry, WB|
|Validated Reactivity||Human, Mouse|
|Cross Reactivity||Predicted to work with mouse, rat and other homologues.|
|Immunogen||A synthetic phospho-peptide corresponding to residues surrounding Tyr774 of human phospho c-Cbl|
|Formulation||1X PBS, 0.02% NaN3, 50% Glycerol, 0.1% BSA|
|Recommended Usage||1µg/mL – 0.001µg/mL. It is recommended that the reagent be titrated for optimal performance for each application. See product image legends for additional information.|
|Pseudonyms||E3 ubiquitin-protein ligase CBL, Casitas B-lineage lymphoma proto-oncogene, Proto-oncogene c-Cbl, RING finger protein 55, RING-type E3 ubiquitin transferase, Signal transduction protein CBL, CBL2, RNF55|
|References||1. Christine, B.F. et al. (2001) Nat. Rev. Mol. Cell Biol. 2: 294-307. 2. Feshchenko, E.A. et al. (1998) J. Biol. Chem. 273: 8323-8331. 3. Blake, T.J. et al. (1991) Oncogene. 6: 653-657. 4. Thien, C.B. and Langdon, W.Y. (1998) Immunol. Cell Biol. 76: 473-482. 5. Kamei, T. et al. (2000) Int. J. Oncol. 17: 335-339.|
Flow cytometric analysis of Daudi cells secondary antibody only negative control (blue) or untreated (red) or treated with IFNα + IL-4 + pervanadate (green) using Phospho-c-Cbl (Tyr774) antibody CblY774-R4C5 at 0.01 µg/mL. Cat. #2361.
Western blot analysis of extracts from Ramos or Jurkat cells, imatinib or pervanadate treated using Phospho-c-Cbl (Tyr774) antibody c-CblY774-R4C5 at 0.1 ug/mL, Cat#2361
Western blot analysis of Jurkat cell extract treated with imatinib or with pervanadate using 0.1 µg/mL Phospho-c-Cbl (Tyr774) antibody CblY774-R4C5 Cat. #2361 or Company A antibody at 0.04 µg/mL (manufacturer’s recommended concentration) developed using the same exposure.