MAP3K3 NanoBRET Kinase Assay

MAP3K3 NanoBRET Target Engagement Intracellular Kinase Assay, competitive binding assay format for screening of compounds binding to the target.

Target

MAP3K3

HGNC Symbol

MAP3K3

Synonyms

MAPKKK3, MEK kinase 3, MEKK3

Cell Line

HEK293 human epithelial cell line from ATCC

Assay Format

384-well plate format using Promega’s NanoBRET TE Intracellular Kinase Assay

Assay Protocol

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.

Readout

Binding of compound to target kinase

Reference Compound IC50s
Compounds IC50 (M)
CEP-701 4.7330e-007
Screening Location

Malvern, USA

Reference compound IC50 for MAP3K3