EGFR [pY845] glyc Antibody | catalog# XBP-4089

Background

Epidermal Growth Factor Receptor, also known as ErbB-1 (EGFR, a 185 kDa glycoprotein) is a transmembrane tyrosine kinase that regulates a variety of biological responses ranging from mitogenesis to stress signaling. The EGFR consists of a large extracellular domain, a single transmembrane domain and a cytoplasmic domain that exhibits kinase activity. Upon binding of EGF to the extracellular domain, the receptor undergoes dimerization and becomes phosphorylated on several tyrosine residues within the cytoplasmic domain. These events result in EGFR activation and increased tyrosine kinase activity toward a variety of intracellular substrates. Tyrosine 845, a major autophosphorylation site in EGFR, is mediated by integrin engagement and Src, and regulates receptor function and tumor progression.

Description

Extracts of A431 cells unstimulated (lane 1) or stimulated with 200 ng/mL EGF for 15 minutes (lanes 2-5) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer overnight at 4˚C, and then incubated with the EGFR [pY845] antibody for two hours at room temperature in a 1% BSA-TBST buffer, following prior incubation with: no peptide (1, 2), the non-phosphopeptide corresponding to the phosphopeptide immunogen (3), a generic phosphotyrosine-containing peptide (4), or the phosphopeptide immunogen (5). After washing, the membrane was incubated with goat F(ab')2 anti-rabbit IgG HRP conjugate and signals were detected using the Pierce SuperSignal™ method.