The Discovery assay enables to determine which of the EPIgeneous binding domain kits must be used for a given epîgenetic binding domain-histone interaction.
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  • Enables fast assay qualification
The Discovery assay enables to determine which of the EPIgeneous binding domain kits must be used for a given epîgenetic binding domain-histone interaction.
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Overview

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.

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.

Validated binding domains

Three kits (A, B and C) have already been validated and fully optimized on a selection of 28 key binding domains. For non-validated reader domains, a fourth one, the Discovery Kit, enables researchers to profile which of the A, B or C kits is the best assay solution.

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