Anti-Phospho-CBL-Y774 Rabbit pAb - P22681

Core Product Specifications and Parameters

Biological Background: CBL Function and Localization

• CBL (casitas B-lineage lymphoma proto-oncogene) is a RING-type E3 ubiquitin-protein ligase that functions as a negative regulator of diverse signaling pathways by mediating ubiquitination of cell surface receptors and associated kinases, leading to their proteasomal degradation (PubMed:10514377, 11896602, 14661060, 14739300, 15190072, 18374639, 21596750). Related references: PMID:10514377 PMID:11896602 PMID:14661060 PMID:14739300 PMID:15190072 PMID:18374639 PMID:21596750

• Specific substrates of CBL include receptor tyrosine kinases (e.g., EGFR, KIT, PDGFR), SRC family kinases, SPRY2, and immune receptors such as LAG3, whose ubiquitination modulates downstream signaling or sorting (PubMed:17094949, 17974561, 40101708, 28381567). Related references: PMID:17094949 PMID:17974561 PMID:40101708 PMID:28381567

• CBL plays a critical role in hematopoietic signaling and is essential for osteoclast differentiation and bone resorption. Phosphorylation at Tyr-731 promotes recruitment of PI3K and is required for osteoclast function (PubMed:14739300). Related references: PMID:14739300

• The protein is localized to the cytoplasm, cell membrane, cilia, and Golgi apparatus, consistent with its roles in endocytic trafficking and signal termination at the plasma membrane.

• CBL contains a conserved N-terminal tyrosine-kinase-binding domain, a RING finger domain responsible for its E3 ligase activity, and a C-terminal proline-rich region with multiple phosphorylation sites, including Tyr-774.

• This antibody specifically recognizes CBL phosphorylated at Tyr-774, a modification that likely influences protein–protein interactions and signal transduction dynamics.

Experimental Guidance and Technical Tips

• The immunogen is a synthetic phosphorylated peptide surrounding Tyr-774; the antibody may preferentially detect activated CBL in signaling-competent samples. For Western blot (WB), consider using phosphatase inhibitors during cell lysis and validating specificity with an unphosphorylated peptide block or dephosphorylation treatment.
• For ELISA, phosphorylated CBL can be captured from cell extracts under non-denaturing conditions; pairing with a total CBL antibody may allow ratiometric phospho‑protein analysis.
• As a rabbit polyclonal antibody, it may exhibit lot‑to‑lot variation; validate each new lot in the relevant experimental system. Store aliquots at -20°C or -80°C to avoid repeated freeze-thaw cycles, and ship on ice to maintain activity.
• No cross-reactivity data with other species is available; for non-human studies, verify reactivity in your model system.

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