ITK NanoBRET Target Engagement Intracellular Kinase Assay for screening of compounds binding to the target.
Target Overview
Target
ITK
HGNC Symbol
ITK
Synonyms
EMT 2; LYK 2; PSCTK2 2
Cell Line
HEK293 human epithelial cell line from ATCC
Assay Properties
Assay Format
384-well plate format using Promega’s NanoBRET TE Intracellular Kinase Assay
Assay Protocol
HEK293 cells transiently expressing NanoLuc- ITK Fusion Vector were seeded into the wells of 384-well plates. The cells were pre-treated with the NanoBRET Tracer K-5 and then treated with reference compound CTx-0294885 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) |
|---|---|
| CTx-0294885 | 1.279e-009 |
Screening Location
Malvern, USA
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