NEK2 NanoBRET Kinase Assay

NEK2 NanoBRET Target Engagement Intracellular Kinase Assay for screening of compounds binding to the target.
Target Overview

Target

NEK2

HGNC Symbol

NEK2

Synonyms

NLK1; HsPK21

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- NEK2 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 1.779e-006

Screening Location

Malvern, USA

Reference compound IC50 for NEK2