MAP3K3 NanoBRET Target Engagement Intracellular Kinase Assay, competitive binding assay format for screening of compounds binding to the target.
MAPKKK3, MEK kinase 3, MEKK3
HEK293 human epithelial cell line from ATCC
384-well plate format using Promega’s NanoBRET TE Intracellular Kinase Assay
HEK293 cells transiently expressing NanoLuc-MAP3K3 Fusion Vector were seeded into the wells of 384-well plates. The cells were pre-treated with the NanoBRET Tracer K-10 and then treated with reference compound CEP-701 for 1 hour. The BRET signal was measured on an Envision 2104 Multilabel Reader. IC50 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