Cellular Phospho-Histone Assay

Cell-based epigenetic assay Phospho-Histone H3 ELISA Assay

Cell Line

HT-29 human colon cell line from ATCC

Assay Format

ELISA

Assay Protocol

HT-29 cells were seeded into the wells of 96-well plate. The cells were pre-treated with reference compound VX-680 (starting at 1 µM, 10-dose with 3-fold dilution) for 90 minutes and then treated with phosphatase inhibitor Calyculinfor 15 minutes. Cells were lysed with cell lysis buffer and cell lysates were collected. Phospho-Histone H3 (serine 10) was measured with ELISA assay 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

Measurement for Phospho-Histone H3 via absorbance

Reference Compound IC50s
Compound IC50 (µM)
VX-680 0.005288

 

Screening location

Malvern, PA, USA

IC50 of HT29 in cell-based phospho-histone assay