Transcreener pADPr PARP FP Assay

The Transcreener^® HTS Assay platform overcomes the need for time-consuming, one-off assay development for individual members within a group transfer enzyme family by utilizing a single set of assay reagents that detect an invariant product. The generic nature of the Transcreener^® HTS Assay platform eliminates delays involved in assay development for new HTS targets, and greatly simplifies compound and inhibitor profiling across multiple target families. Transcreener assays have been validated with major HTS instruments to determine the optimal filters and settings for maximal assay performance. Application notes with all important details are available.

The Transcreener® pADPr PARP FP 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, competitive fluorescence polarization (FP) assay. As an example, the Transcreener pADPr PARP FP 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 FP 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 10 nM to 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.