Cell-based epigenetic assay Phospho-Histone H3 ELISA Assay
HT-29 human colon cell line from ATCC
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 Calyculin A for 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.
Measurement for Phospho-Histone H3 via absorbance
Malvern, PA, USA