Cellular BRD4 Protein Degradation Assay

Cellular BRD4 Protein Degradation in PROTAC dBET6 Treated HepG2 Cells by Western Blot

Target Overview



HGNC Symbol




Cell Line

HepG2 cells from ATCC

Assay Properties

Assay Format

Western Blot

Assay Protocol

HepG2 hepatocellular cancer cells were seeded into the wells of 12-well plates. The cells were treated with reference PROTAC compound dBET6 for 8 hours. The cell lysates were subjected to Western blot analysis. The blots were probed with BRD4 antibody and α-Tubulin antibody. The membrane was scanned with LI-COR Odyssey Fc Imaging System. The specific bands of interest were quantified by LI-COR Image Studio Lite software. The IC50 curve was plotted and IC50 value was calculated using the GraphPad Prism program based on a sigmoidal dose-response equation.


LI-COR Odyssey Fc Imaging System

Reference Compound IC50s
Compounds IC50 (M)
dBET6 2.332e-008

Screening Location

Malvern, PA, USA

Further information

More information can be found on our website Protein Degradation Assay – PROTAC Screening

Western blot for detection of BRD4 degradation in PROTAC dBET6 treated HepG2 cells

Western blot for detection of BRD4

Western blot quantification for IC50 determination of PROTAC dBET6 induced BRD4 degradation

IC50 determination of BRD4 degradation