The epidermal growth factor receptor (EGFR; ErbB-1; HER1 in humans) is the cell-surface receptor for members of the epidermal growth factor family (EGF-family) of extracellular protein ligands. The epidermal growth factor receptor is a member of the ErbB family of receptors, a subfamily of four closely related receptor tyrosine kinases: EGFR (ErbB-1), HER2/c-neu (ErbB-2), Her 3 (ErbB-3) and Her 4 (ErbB-4). Mutations affecting EGFR expression or activity could result in cancer.
Binding of EGF leads to autophosphorylation of the EGF Receptor at 5 tyrosine residues. In addition, EGF Receptor is phosphorylated by members of the src kinase family at different tyrosine residues, phosphotyrosine 845 in the activation loop being the major site (tyrosine 845 is not an autophosphorylation site). EGFR acquires its full kinase activity only upon autophosphorylation and phosphorylation at tyrosine 845. E3375-53F allows the detection of EGFR activation after interaction with src kinases. Specifically recognizes EGFR phosphorylated at Tyrosine 845. The antibody does not interact with the non-phosphorylated EGFR nor with unrelated Tyrosine-phosphorylated proteins.
Applications:
Suitable for use in ELISA, Western Blot.. Other applications not tested.
Recommended Dilution:
ELISA: 0.1ug/ml
Western Blot: 0.1ug/ml for HRP/ECL detection.
Optimal dilutions to be determined by the researcher.
Positive Control:
Cell lysate of 2 x 10e6 vanadate treated HepG2 cells. Supplied as a lyophilized powder. Reconstitute with 200ul ddH2O. After complete solubilization of the proteins, add 200ul SDS-PAGE sample buffer and incubate at 90 degrees C for 5 minutes.
Western Blot : 20ul/lane HRP/ECL
Storage and Stability:
Lyophilized powder may be stored at -20 degrees C. Reconstitute with sterile 40-50% glycerol, aliquot and store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.