Transcreener(R) pADPr PARP TR-FRET Assay

The Transcreener(R) pADPr PARP TR-FRET Assay is a biochemical HTS assay for measuring the production of poly(ADP-ribose) (pADPr) in poly(ADP-ribose) polymerase (PARP) reactions. The assay relies on pADPr Coupling Enzymes (CE) to convert pADPr into AMP, which is then detected using a far-red, timeresolved Förster-resonance-energy-transfer (TR-FRET) readout. As an example, the Transcreener pADPr PARP TR-FRET assay can be used to detect the activity of human PARP1, which uses NAD+ to form pADPr on itself, histones, and DNA repair proteins. The Transcreener pADPr PARP TR-FRET assay is designed specifically for high throughput screening (HTS), with a single-addition, mix-and-read format. It is easy to integrate into automated HTS workflows, with outstanding reagent stability (deck stability > 16 hours, signal stability > 24 hours), robust detection of ADPR monomers released from pADPr over a range of down to 10 nM and as high as 100 µM, and compatibility with commonly used multimode plate readers. Data quality is excellent (Z' > 0.7), and the assay uses a far-red tracer to minimize interference from fluorescent compounds and light scattering.