Purification notesab74091 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/500 - 1/1000. Detects a band of approximately >117 kDa (predicted molecular weight: 114 kDa).
1/50 - 1/100.
1/500 - 1/1000.
FunctionNon-receptor tyrosine kinase that regulates the activity of a number of proteins by tyrosine phosphorylation especially proteins critical for cell survival, cell growth, and proliferation. Activates AKT1 by phosphorylating it on 'Tyr-176' resulting in its activation. Phosphorylates AR on 'Tyr-267' and 'Tyr-363' and promotes its recruitment to the androgen-responsive enhancers (AREs). Phosphorylates WWOX on 'Tyr-287'. Downstream effector of CDC42 which mediates CDC42-dependent cell migration via phosphorylation of BCAR1. Binds to both poly- and mono-ubiquitin and regulates ligand-induced degradation of EGFR. Participates in clathrin-mediated endocytosis. May be involved both in adult synaptic function and plasticity and in brain development.
Tissue specificityThe Tyr-284 phosphorylated form shows a significant increase in expression in breast cancers during the progressive stages i.e. normal to hyperplasia (ADH), ductal carcinoma in situ (DCIS), invasive ductal carcinoma (IDC) and lymph node metastatic (LNMM) stages. It also shows a significant increase in expression in prostate cancers during the progressive stages.
Sequence similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family. Contains 1 CRIB domain. Contains 1 protein kinase domain. Contains 1 SH3 domain. Contains 1 UBA domain.
DomainThe EBD (EGFR-binding domain) domain is necessary for interaction with EGFR. The SAM-like domain is necessary for NEDD4-mediated ubiquitination. Promotes membrane localization and dimerization to allow for autophosphorylation. The UBA domain binds both poly- and mono-ubiquitin.
Post-translational modificationsAutophosphorylation regulates kinase activity. Phosphorylation on Tyr-518 is required for interaction with SRC. Ubiquitinated by NEDD4. Its EGF-induced degradation is EGFR activation-dependent and is processed by lysosomes, not proteasomes.
Cellular localizationCell membrane. Nucleus. Endosome. Cell junction > adherens junction. Cytoplasmic vesicle membrane. The Tyr-284 phosphorylated form is expressed both in the membrane and nucleus. Co-localizes with EGFR on the endosomes.