武漢云克隆科技股份有限公司

The epidermal growth factor receptor (EGFR) is a growth factor receptor that induces cell differentiation and proliferation upon activation through the binding of one of its ligands. The receptor is located at the cell surface, where the binding of a ligand activates a tyrosine kinase in the intracellular region of the receptor. This tyrosine kinase phosphorylates a number of intracellular substrates that activates pathways leading to cell growth, DNA synthesis and the expression of oncogenes such as fos and jun. Epidermal growth factor (EGF) family peptides are ligands for the EGF receptor (EGFR), thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human EGFR and recombinant human EGF. Briefly, biotin-linked EGFR were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to EGF-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with Streptavidin-HRP for 30min, then wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 μl stop solution to the wells and read at 450 nm immediately. The binding activity of recombinant human EGFR and recombinant human EGF was shown in Figure 1, the EC50?for this effect is 11.46 ug/mL.