FLT3 (D835Y) NanoBRET Target Engagement Intracellular Kinase Assay for screening of compounds binding to the target.
Target Overview
Target
FLT3 (D835Y)
HGNC Symbol
FLT3 (D835Y)
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 (D835Y) 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): 4.373e-009 M
Screening Location
Malvern, USA
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