Synthetic phosphopeptide corresponding to residues surrounding Tyr774 of human CBL.
Species Reactivity
Human
Validated Applications
WB
Target Molecular Weight
Predicted band size: 100 kDa
Positive Control
A549 starved for 2 hours then add 1μM sodium orthovanadate for 1 hour cell lysate, HeLa treated with 1mM sodium orthovanadate for 30 minutes cell lysate.
Conjugation
unconjugated
Clone Number
JE62-93
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Lot Concentration Lookup
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Cbl (named after Casitas B-lineage Lymphoma) is a mammalian gene family. CBL gene, a part of the Cbl family, encodes the protein CBL which is an E3 ubiquitin-protein ligase involved in cell signalling and protein ubiquitination. Mutations to this gene have been implicated in a number of human cancers, particularly acute myeloid leukaemia. Ubiquitination is the process of chemically attaching ubiquitin monomers to a protein, thereby targeting it for degradation. As this is a multi-step process, several different enzymes are involved, the final one being a member of the E3 family of ligases. Cbl functions as an E3 ligase, and therefore is able to catalyse the formation of a covalent bond between ubiquitin and Cbl's protein substrate - typically a receptor tyrosine kinase. The RING-finger domain mediates this transfer, however like other E3 ligases of the RING type no intermediate covalent bond is formed between ubiquitin and the RING-finger domain. The stepwise attachment of ubiquitin to the substrate receptor tyrosine kinase can lead to its removal from the plasma membrane and subsequent trafficking to the lysosome for degradation.
Background References
1. Leardini D et al. Role of CBL Mutations in Cancer and Non-Malignant Phenotype. Cancers (Basel). 2022 Feb
2. Pinilla-Macua I et al. Cbl and Cbl-b independently regulate EGFR through distinct receptor interaction modes. Mol Biol Cell. 2023 Dec
Western blot analysis of Phospho-CBL (Y774) on different lysates with Rabbit anti-Phospho-CBL (Y774) antibody (HA722786) at 1/1,000 dilution.
Lane 1: A549 cell lysate Lane 2: A549 starved for 2 hours cell lysate Lane 3: A549 starved for 2 hours then add 1μM sodium orthovanadate for 1 hour cell lysate Lane 4: HeLa cell lysate Lane 5: HeLa treated with 1mM sodium orthovanadate for 30 minutes cell lysate Lane 6: A549 starved for 2 hours then add 1μM sodium orthovanadate for 1 hour cell lysate, then the membrane treated with λpp for 1 hour Lane 7: HeLa treated with 1mM sodium orthovanadate for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour
Lysates/proteins at 10 µg/Lane.
Predicted band size: 100 kDa Observed band size: 100-110 kDa
Exposure time: 59 seconds; ECL: K1802;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722786) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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