HEK293 human epithelial cell line from ATCC
384-well plate format
HEK293 cells transiently expressing IRAK3-NanoLuc fusion vector were seeded into the wells of 384-well plates. The cells were pre-treated with the tracer 05 and then treated with reference compound CTx-0294885 and Crizotinib for 1 hour. NanoBRET signal was measured on an Envision 2104 Multilabel Reader. IC50 (EC50) value was calculated and IC50 curve was plotted using the GraphPad Prism 4 program based on a sigmoidal dose-response equation.
Binding of compound to target kinase