AKT1 NanoBRET Kinase Assay

AKT1 NanoBRET Target Engagement Intracellular Kinase Assay for screening of compounds binding to the target.

Target Overview

Target

AKT1

HGNC Symbol

AKT1

Synonyms

PKB; RAC; PRKBA; RAc-ALPHA

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- AKT1 Fusion Vector were seeded into the wells of 384-well plates. The cells were pre-treated with the NanoBRET Tracer K-10 and then treated with reference compound GSK690693 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 program based on a sigmoidal dose response equation.

Readout

Binding of compound to target kinase

Reference Compound IC50s
Compounds IC50 (M)
GSK690693 2.360e-007

Screening Location

Malvern, PA, USA

Reference Compound IC50 for AKT1

Reference Compound IC50 for AKT1

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