Product Name:
CBL-pY674
Product Number:
ab-pn743
Target Full Name: E3 ubiquitin-protein ligase CBL
Target Alias: Cas-Br-M (murine) ecotropic retroviral transforming; Cas-Br-M (murine) ecotropic retroviral transforming sequence; CBL E3 ubiquitin protein ligase; CBL2; Oncogene CBL2; Proto-oncogene c-CBL; RNF55; Signal transduction protein CBL
Product Type Specific: Cbl phosphosite-specific antibody
Antibody Code: PN743
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: Y674
Protein UniProt: P22681
Protein SigNET: CBL
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Synthetic phosphopeptide patterned after human CBL
Antibody Immunogen Sequence: NAI(pY)SLAAC
Antibody Immunogen Description: Corresponds to amino acid residues N671 to A678. The effect of Y674 phosphorylation is unclear. This is the major in vivo phosphorylation site in Cbl (≥732 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus). This human phosphosite is highly conserved in vertebrates and also found in fish.
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 0.2 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Chimpanzee | Rhesus macaque | Mouse | Chicken | Zebra fish.
Scientific Background: CBL (E3 ubiquitin-protein ligase CBL) is an adaptor/scaffold and E3 ubiquitin-protein ligase that acts as a negative regulator of many signaling pathways by mediating ubiquitination of cell surface receptors. It transfers ubiquitin from specific E2 ubiquitin-conjugating enzymes to transfers it to substrates promoting their degradation by the proteasome. Among the activated receptor-tyrosine kinases that binding CBL are KIT, FLT1, FGFR1, FGFR2, PDGFRA, PDGFRB, CSF1R, EPHA8 and KDR, and it mediates their ubiquitination to terminate signalling. It also binds membrane-bound HCK, SRC and other kinases of the SRC family and mediates their ubiquitination and degradation. It is involved in LAG3-mediated inhibition of TCR signaling: following ligand-binding to LAG3, by catalyzes 'Lys-63'-linked ubiquitination of LAG3, unleashing the LAG3 C-terminus from the membrane, and initiating a signaling that prevents TCR activation. It ubiquitinates NECTIN1 following association between NECTIN1 and herpes simplex virus 1/HHV-1 envelope glycoprotein D, leading to NECTIN1 removal from cell surface. CBL participates in signal transduction in hematopoietic cells, and regulates osteoblast differentiation and apoptosis, and is essential for osteoclastic bone resorption. The 'Tyr-731' phosphorylated form of CBL induces the activation and recruitment of phosphatidylinositol 3-kinase to the cell membrane in a signaling pathway that is critical for osteoclast function. CBL proteins contain a characteristic N-terminal structure, including a Tyrosine Kinase-Binding (TKB) Domain (binds to specific tyrosine-phosphorylated targets); a Linker Region (connects the TKB and RING domains); a RING Finger Domain (essential for transferring ubiquitin from E2 enzymes to the substrate); and Proline-rich and UBA domains (enable interaction with SH3-domain-containing proteins and ubiquitin, respectively). Mutations or dysregulation of CBL genes are implicated in various cancers, including acute myeloid leukaemia (AML) and myeloproliferative disorders. This description may include information annotated by UniProt and/or Google AI.