Enzolution™ PARP1 FP Assay System

The Enzolution™ PARP1 Assay System is intended for use with the Transcreener® pADPr PARP Assay Kits (Parts #3043 and #3044) to measure the enzymatic activity of PARP1 (poly(ADP-ribose) polymerase 1). PARP1 forms poly(ADP-ribose) (pADPr) chains on target proteins, including itself, using NAD+ as a donor substrate (Figure 1). The Transcreener pADPr PARP Assay Kits use pADPr Coupling Enzymes (CE) to convert pADPr formed by PARP1 to AMP, which is then detected using far-red, competitive fluorescence assays that enable single addition, mix-and-read detection in continuous or endpoint formats. 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 PARP1 Assay System provides all reagents required to screen and profile PARP1 inhibitors when used with the Transcreener pADPr PARP Assay Kits, including purified, full length human PARP1 (aa 1-1014) and Sheared Salmon Sperm DNA. Note that PARP1 can ADP-ribosylate multiple substrates, including itself. The assay was optimized to detect the auto-ADP-ribosylation reaction, so it is not necessary to add any other protein substrate. The protocol is configured for 384-well plates; use of different multi-well plate formats will require adjustment of reagents concentrations utilized in the assay.

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