PARPtrap™ Assay Kit

Catalog #: 80584
Size: 96 reactions
$1,095
Background:
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.
Description:

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 (Fig. 1).

PARPtrap Assay Kit

Figure 1. PARPtrap™ Assay Kit schematic

Synonym(s):
poly (ADP-ribose) polymerase 1, poly(ADP-ribosyl)transferase, ADPRT, PARP1
Format:

COMPONENTS:

Instructions for use:
See assay kit datasheet for detailed protocol.
Storage / Stability:
Up to 1 year when stored as recommended.
Application(s):
Great for screening small molecules that enhance PARP1/DNA trapping for drug discovery and HTS applications.
Reference(s):

1. Murai, J., et al. Molecular Cancer Therapeutics 2014. 13:433-443.
2. Murai, J., et. al. Cancer Research 2012. 72:5588-5599.

Notes:

MATERIALS OR INSTRUMENTS REQUIRED BUT NOT SUPPLIED:
Fluorescent microplate reader capable of measuring fluorescence polarization
Adjustable micropipettor and sterile tips
Rotating or rocker platform

Warning(s):
Avoid freeze/thaw cycles.
Scientific Category:
PARP
Product Type:
Assay Kit
Data shown is lot-specific. Contact us for specific information on other lots.

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Product Datasheets

80584.pdf