PARP1 is known to bind damaged DNA through its N-terminal zinc finger domain. After PARP1 ribosylates itself (autoribosylation), it dissociates from DNA due to the accumulated negative charge of the ribosyl polymer. Trapped PARP-DNA complexes have been shown to be cytotoxic to cancer cells.
The PARPtrap Assay Kit is designed to measure PARP1/DNA complex formation in a high throughput screening assay using fluorescence polarization (FP). The PARPtrap Assay Kit comes in a convenient 96-well format, with purified PARP1 enzyme, fluorescent labeled nicked DNA, and PARPtrap assay buffer for 100 enzyme reactions. The key to the PARPtrap Assay Kit is the fluorescent labeled nicked DNA. Without the PAR reaction, PARP1 binds to the fluorescent labeled nicked DNA, resulting in the emission of highly polarized light. However, after autoribosylation of PARP1, the nicked DNA is dissociated from PARP1 and rotates freely, emitting less polarized light.
Great for screening small molecules that enhance PARP1/DNA trapping for drug discovery and HTS applications.
Up to 1 year when stored as recommended.
PARP1, GST-tag: 5 µg; -80°C 50x Fluorescent labeled nicked DNA: 0.1 ml; -80°C 5x PARPtrap assay buffer: 1.5 ml; -80°C 10x NAD+: 0.5 ml; -80°C Black 96-well plate: 1; Room Temp. Plate lid: 1; Room Temp.