Shopping Cart » DIVAATM     » Cell Invasion     » PathClear® BME     » PARP     » CometAssayTM     » HT 8-oxo-dG ELISA  Home » DNA Damage DNA Damage HT Fluorescent Homogeneous PARP Inhibition Assay Trevigen's Homogeneous PARP Inhibition Assay is a highly sensitive fluorescent screening assay for the rapid identification of PARP-1 inhibitors in an in vitro system. This one hour endpoint assay is performed in two successive steps requiring only the consecutive addition of reaction components. A PARP reaction is first performed followed by a detection step. Inhibitors are identified by an increase in fluorescent signal when PARP mediated NAD+ depletion is inhibited. The level of NAD+ is coupled to a cycling assay involving alcohol dehydrogenase and diaphorase. Each time NAD+ cycles through these coupled reactions, a molecule of highly fluorescent resorufin is generated. Alcohol dehydrogenase (AD) reduces NAD+ to NADH, while the diaphorase cycles NADH back to NAD+ with the generation of a highly fluorescent resorufin molecule (from the non-fluorescent substrate, resazurin). The number of cycles can be controlled by the time of incubation to adjust assay sensitivity, as needed, and the reaction is terminated by the addition of a stop solution. In addition, the assay can be used to determine relative IC50 values for PARP inhibitors and is capable of detecting as little as 10% inhibition of PARP-1 activity. The Homogeneous PARP Inhibition Assay is designed for the screening of PARP inhibitors using a NAD concentration below the Km of PARP. Once potential inhibitors are identified, results can be confirmed using Trevigen's HT Universal PARP Assay kits (cat# 4676-096-K or 4677-096-K). Features: Fluorescent, non-radioactive format High throughput 96 test size Detects as little as 10% inhibition of PARP-1 activity   Applications: Identification of PARP-1 inhibitors   Components: Part # Component Size   4690-096-01 10X Cycling Enzymes 600 µl   4690-096-02 200 nM NAD 3.5 ml   4690-096-03 PARP HSA Enzyme 200 ng/ml 100 µl   4690-096-04 Activated DNA, 200 ng/µl 600 µl   4690-096-05 Resazurin 600 µl   4690-096-06 10X Buffer (1M Tris-HCl, pH 8) 3.5 ml   4690-096-07 Stop Solution 6 ml   Storage: Components are stored at -20°C and Room Temperature   References: 1. Meyer-Ficca, M.L., et al., Poly(ADP-ribose) polymerases: managing genome stability. Int J Biochem Cell Biol, 2005. 37(5): p. 920-6. 2. Hassa, P.O., et al., Nuclear ADP-ribosylation reactions in mammalian cells: where are we today and where are we going? Microbiol Mol Biol Rev, 2006. 70(3): p. 789-829. 3. Virag, L. and C. Szabo, The therapeutic potential of poly(ADP-ribose) polymerase inhibitors. Pharmacol Rev, 2002. 54(3): p. 375-429. Top   Ordering Information Catalog #: Price: 4690-096-K HT F Homogeneous PARP Inhibition Assay, 96 Tests $450.00 Trevigen Products are now available for ordering through Download Protocol Download Worksheet Related Products HT Chemiluminescent PARP/Apoptosis Assay HT Colorimetric PARP/Apoptosis Assay HT Chemiluminescent PARG Assay Kit HT Colorimetric PARG Assay Kit Universal Colorimetric PARP Assay Kit Universal Chemiluminescent PARP Assay Kit PARP Inhibitors TREVIGEN INC. :: 8405 HELGERMAN CT. :: GAITHERSBURG, MD 20877 1.800.TREVIGEN :: 1.800.873.8443 :: (FAX) 1.301.560.4973 :: INFO@TREVIGEN.COM