PIK3CA (H1047Y)/PIK3R1 NanoBRET Kinase Assay

PIK3CA (H1047Y)/PIK3R1 NanoBRET Target Engagement Intracellular Kinase Assay for screening of compounds binding to the target.

Target Overview

Target

PIK3CA (H1047Y)/PIK3R1

HGNC Symbol

PIK3CA (H1047Y)-PIK3R1

Synonyms

MGC142161, MGC142163, p110-alpha, PI3K, PI3K-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 co- expressing NanoLuc- PIK3CA(H1047Y) Fusion Vector and PIK3R1 Expression Vector were seeded into the wells of 384-well plates. The cells were pre-treated with the NanoBRET Tracer K-3 and then treated with reference compound PI-103 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)
PI-103	1.971e-008

Screening Location

Malvern, PA, USA

Further Information

More information can be found on our website NanoBRET Target Engagement Intracellular Kinase Assay Services

Reference compound IC50 for PIK3CA (H1047Y)/PIK3R1

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