FLT3 (D835V)

NanoBRET Target Engagement Intracellular Kinase Assay, Competitive binding assay format

Target

FLT3 (D835V)

HGNC Symbol

FLT3 (D835V)

Synonyms

FLK2; STK1; CD135

Cell Line

HEK293 human epithelial cell line from ATCC

Assay Format

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

Assay Protocol

HEK293 cells transiently expressing NanoLuc®- FLT3 (D835V) Fusion Vector were seeded into the wells of 384-well plates. The cells were pre-treated with the NanoBRETTM Tracer K-10 and then treated with reference compound Foretinib 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

Foretinib (IC50 value): 4.273e-009 M

Screening Location

Malvern, USA

Reference Compound IC50 for FLT3 (D835V)