ERBB2 Cellular Phosphorylation Assay (intracellular kinase activity assay) for compound screening and profiling in intact cells
The receptor tyrosine kinase ERBB2 belongs together with EGF-R and ERBB3/B4 to the ERBB receptor family. These receptors transduce a variety of signals crucial for the processes of cellular proliferation, survival and apoptosis. Since ERBB2 lacks a physiological ligand, the activation mechanism involves either homo- or heterodimerization with other ligand-activated ERBB receptors. Importantly, overexpression of ERBB2 was found to be responsible for cellular transformation and could be identified in several cancer types.
NEU, NGL, HER2, TKR1
Reaction Biology’s cellular ERBB2 phosphorylation assay was generated based on a NIH3T3 background. Cells were transfected to express a full-length ERBB2. After clonal selection a transformed cell line with a high level of autophosphorylated ERBB2 was obtained. By adding Lapatinib phospho-ERBB2 levels are largely decreased and thus the dynamic behaviour to determine inhibitory potentials of compounds was achieved. Phospho-ERBB2 levels are quantified by Sandwich-ELISA technique. The assay is validated based on known inhibitors of ERBB2 kinase activity (see Fig. 1).
Substrate phosphorylation as a readout of intracellular kinase activity via ELISA
More information can be found on our website Cellular Phosphorylation Assay Services.
Reference compound IC50 for ERBB2
Lapatinib is a potent inhibitor of the phosphoERBB2 signal found in the described cells. The graph shows representative results.