The PARP1 Colorimetric Assay Kit is designed to measure PARP1 activity for screening and profiling applications. PARP1 is known to catalyze the NAD-dependent addition of poly(ADP-ribose) to histones. The key to the PARP1 Colorimetric Assay Kit is the biotinylated NAD+. With this kit, only three simple steps are required for PARP1 reactions. First, histone proteins are coated on a 96-well plate. Next, a biotinylated NAD+ mix (termed PARP Substrate Mixture) is incubated with the PARP1 enzyme and an activated DNA template in an optimized assay buffer. Finally, the plate is treated with streptavidin-HRP followed by addition of the colorimetric HRP substrate to produce color that can be measured using a UV/Vis spectrophotometer microplate reader.

Figure 1. PARP1 Colorimetric Assay Kit schematic

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

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