The PARP14 Chemiluminescent Assay Kit is designed to measure PARP14 activity for screening and profiling applications. PARP14 is known to catalyze the NAD-dependent ADP-ribosylation of histones. The PARP14 assay kit comes in a convenient 96-well format, with purified PARP14 enzyme, histone mixture, and PARP assay buffer for 100 enzyme reactions. The key to the PARP14 Chemiluminescent Assay Kit is the biotinylated NAD+. With this kit, only three simple steps are required for PARP14 reactions. First, histone proteins are coated on a 96-well plate. Next, a biotinylated NAD+ mix (termed PARP Substrate Mixture) is incubated with the PARP14 enzyme in an optimized assay buffer. Finally, the plate is treated with streptavidin-HRP followed by addition of the ELISA ECL substrate to produce chemiluminescence that can be measured using a chemiluminescence reader.

Figure 1. PARP14 Chemiluminescent Assay Kit schematic

*NOTE: As of April 2022, this protocol has been re-optimized for performance. Previous versions of this kit are available upon request.