Revvity, 62BDDPEG, HTRF EPIgeneous Binding Domain Discovery Kit, 500 Assay Points

The EPIgeneous Binding Domain kit series provides a simple biochemical approach to study epigenetic reader interactions with modified histones. The three kits, A, B and C, have been validated on 28 different bromo-, chromo- and tudordomains. All kits are based on a GST-tagged binding domain /biotin-coupled Histone peptide format, and can be run using the same add-and-read single plate protocol. Whenever a given domain is not part of the list of validated interactions, the Discovery Binding Domain kit enables researchers to profile which of the A, B or C kits is the best assay solution. HTRF assays offer many advantages over other technologies: Homogeneous add-and-read format No wash steps Low background Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates Stable signal, providing flexibility in time of readout or size of assays
Assay principle In EPIgeneous Binding Domain kits, the GST-tagged reader domain protein binding to the biotinylated peptide substrate is detected by a conjugate mix: anti-GST Eu or Tb cryptate-labeled antibody conjugate (donor), and streptavidin conjugated to either d2 or XL665 (acceptor). The interaction of the reader domain with the substrate brings the donor and acceptor dyes into close proximity, and allows FRET to occur upon light excitation. The specific signal at 665 nm is inhibited when a specific compound prevents the reader domain protein from binding to its substrate.