AlphaLISA BRD4 (BD1) and H4 K12Ac Binding Assay Kit
This kit is designed for the detection of binding activity BRD4 BD1 and acetylated histone H4 peptide (H4 K12Ac) using a homogeneous AlphaLISA assay - no wash steps. This assay allows screening of BRD4 inhibitors with a highly-specific substrate.
AlphaLISA® detection of epigenetic marks. Glutathione (GSH) AlphaLISA Acceptor beads are used to capture the GST-tagged bromodomains and Streptavidin-coated Donor beads capture the biotinylated peptide. Donor and Acceptor beads come into proximity through BRD4 binding to the acetylated histone H4 peptide. Excitation of the Donor beads (680 nm) provokes the release of a singlet oxygen that triggers a cascade of energy transfer reactions in the Acceptor beads, resulting in a sharp peak of light emission at 615 nm.
Bromodomains (BRDs) are a class of epigenetic reader molecules that translate the acetylation state of histones into downstream cell-signaling events. Due to their importance in these physiological processes, bromodomains are believed to be involved in many disease states, including cancer and atherosclerosis. BRD4 directly maintains the acetylation state of histones and preferentially binds to acetylated H3 and H4. This assay can be used to determine the binding activity of BRD4 with acetylated histone H4 peptide and to measure the inhibition of this binding.