The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 114 kDa).
Non-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.
The 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.
Belongs 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.
The 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.
Autophosphorylation 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.
Cell 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.
ICC/IF image of ab65108 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab65108, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Maxson JE et al. Identification and Characterization of Tyrosine Kinase Nonreceptor 2 Mutations in Leukemia through Integration of Kinase Inhibitor Screening and Genomic Analysis. Cancer Res76:127-38 (2016).
Read more (PubMed: 26677978) »
Wilson C et al. Overcoming EMT-associated resistance to anti-cancer drugs via Src/FAK pathway inhibition. Oncotarget5:7328-41 (2014).
Read more (PubMed: 25193862) »