The TNKS2 Colorimetric Assay Kit is designed to measure the activity of Tankyrase 2 (TNKS2, also known as PARP5B) for screening and profiling applications. TNKS2 is known to catalyze the NAD-dependent ADP-ribosylation of histones. The key to the TNKS2 Colorimetric Assay Kit is the biotinylated NAD+. First, histone proteins are coated on a 96-well plate. Next, a biotinylated NAD+ mix (termed PARP Substrate Mixture) is incubated with the TNKS2 enzyme. 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. TNKS2 (PARP5B) Colorimetric Assay Kit schematic

*NOTE: As of March 2023, this protocol has been re-optimized for performance. Previous versions of this kit are available upon request.