Cell-based epigenetic assay for the detection of HDAC activity in cancer cells via quantification of acetylated tubulin.
Western Blot
U266B1 cells (source: ATCC) are seeded into the wells of 24-well plates. The cells are treated with reference compound TSA (Trichostatin A) for 16 hours. The cell lysates are subjected to Western blot analysis. The blots are probed with Acetylated Tubulin antibody and then reprobed with Tubulin antibody. The membrane is scanned with LI-COR Odyssey Fc Imaging System. The specific bands of interest are quantified by LI-COR Image Studio Lite software. The IC50 curve is plotted and IC50 value is calculated using the GraphPad Prism program based on a sigmoidal dose-response equation.
LI-COR Odyssey Fc Imaging System
| Compound | IC50 (µM) |
|---|---|
| TSA | 0.750 |
Malvern, PA, USA
Western blotting for detection of acetylated tubulin in Trichostatin A treated cells.
Western blot quantification performed with the LI-COR Odyssey Fc Imaging System for IC50 determination of Trichostastin A.
