ERBB4 Cellular Phosphorylation Assay (intracellular kinase activity assay) for compound screening and profiling in intact cells
The receptor tyrosine kinase ERBB4 (Erythroblastic Leukemia Viral Oncogene Homolog-4) belongs together with EGF-R, ERBB2 and ERBB3 to the ERBB receptor family. It binds members of the neuregulin and the EGF family (NRG1-4, BTC, EREG and HBEGF). Binding of ligand results in receptor homo- or heterodimerization with EGF or ERBB2 and subsequently receptor autophosphorylation. ERBB4 signaling regulates cell proliferation, differentiation, migration and apoptosis. It plays an important role in the development of the heart, the central nervous system and the mammary gland.
ERBB4
HER4
T47D
Endogenous
The human breast cancer cell line T47D is known to endogenously express the receptor tyrosine kinase ERBB4. Stimulation of these cells with the physiological ligand Neuregulin results in a robust receptor autophosphorylation. Compounds are preincubated before cell stimulation to allow thorough target binding. Stimulation conditions are optimized to determine dose-related inhibition of the phospho-ERBB4 signal, which is subsequently quantified by Sandwich-ELISA technique. Figure 1 shows data confirming the inhibitory activity of Lapatinib, a cognate ERBB4 inhibitor.
Substrate phosphorylation as a readout of intracellular kinase activity via ELISA
Freiburg, Germany
More information can be found on our website Cellular Phosphorylation Assay Services.
Reference compound IC50 for ERBB4
Lapatinib is a potent inhibitor of the phosphoERBB4 signal as observed in T47D breast cancer cells. The graph shows a representative result.