Enzolution™ PARP2 TR-FRET Assay System

The Enzolution™ PARP2 Assay System is intended for use with the Transcreener® pADPr PARP Assay Kits (Parts #3043 and #3044) to measure the enzymatic activity of PARP2 (poly(ADP-ribose) polymerase 2). Upon binding to DNA, PARP2 forms poly(ADP-ribose) (pADPr) chains on target proteins, including itself, using NAD+ as a donor substrate (Figure 1). The assay relies on Coupling Enzymes (CE) to convert pADPr into AMP, which is detected using a far-red, fluorescence polarization (FP) or time-resolved Förster-resonance-energy-transfer (TR-FRET) assay. The assay has been optimized and extensively validated for high-throughput screening (HTS) and inhibitor dose response measurements using most multimode plate readers.

The Enzolution PARP2 Assay System provides all reagents required to screen and profile PARP2 inhibitors when used with the Transcreener pADPr PARP Assay Kits, including purified, full-length human PARP2 and sheared salmon sperm DNA. Since PARP2 can add pADPr to itself, this assay was optimized to detect the auto-ADP-ribosylation reaction and does not require the addition of other protein substrates. The protocol is configured for 384-well plates; use of different multi-well plate formats will require adjustment of reagent concentrations utilized in the assay.

When purchasing this Enzolution Assay System, it is important to choose the assay system readout that corresponds to the TR-FRET configuration of your Transcreener pADPr Assay.