FLT3 (D835H) NanoBRET Kinase Assay

FLT3 (D835H) NanoBRET Target Engagement Intracellular Kinase Assay for screening of compounds binding to the target.
Target Overview

Target

FLT3 (D835H)

HGNC Symbol

FLT3 (D835H)

Synonyms

FLK2; STK1; CD135

Cell Line

HEK293 human epithelial cell line from ATCC

Assay Properties

Assay Format

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

Assay Protocol

HEK293 cells transiently expressing NanoLuc®- FLT3 (D835H) 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): 2.547e-009 M

Screening Location

Malvern, USA

Reference Compound IC50 for FLT3 (D835H)