PARPtrap™ Assay Kit for PARP2
The PARPtrap™ Assay Kit for PARP2 is designed to measure PARP2/DNA complex formation in a high throughput screening assay using fluorescence polarization (FP). Similarly, to PARP1, PARP2 recognizes and binds damaged DNA via its DNA-binding domain. Binding to DNA activates PARP2 and in the presence of NAD+ PARP2 ribosylates itself (auto-ribosylation), which leads to PARP2 dissociation from the DNA due to the accumulated negative charge of the ribosyl polymer. In the presence of some inhibitors, however, PARP remains bound to the DNA, a phenomenon termed trapping. Trapped PARP-DNA complexes have been shown to be highly cytotoxic to cancer cells, therefore such inhibitors may be desirable for cancer treatment.
The key to the PARPtrap™ Assay Kit is the fluorescent-labeled oligonucleotide duplex. In the absence of ribosylation, PARP2 binds to the fluorescent probe, forming a large complex and resulting in the emission of highly polarized light. However, after auto-ribosylation PARP2 dissociates from the oligonucleotide duplex, which then rotates freely, emitting less polarized light (Fig. 1). Addition of a PARP2 inhibitor results in trapping of PARP2 to the fluorescent oligonucleotide duplex, and increases the FP signal in a dose dependent manner.
The PARPtrap™ Assay Kit is a fluorescence polarization homogeneous assay. The FP signal is measured using a fluorescent microplate reader capable of measuring fluorescence polarization.
Figure 1. PARPtrap™ Assay Kit for PARP2 schematic
Need us to run inhibitor screens or profile your compounds against PARPtrap™ for PARP2? Check out our PARP/PARPTrap™ Screening Services or DNA Replication and Repair Screening Services.
- DTT
- Fluorescent microplate reader capable of measuring fluorescence polarization
- Adjustable micropipettor and sterile tips
- Rotating or rocker platform
96 Reactions
| Catalog # | Name | Amount | Storage |
| 80502 | PARP2, GST-tag* | 8 µg | -80°C |
| 78297 | Fluorescent labeled nicked oligonucleotide duplex (100 nM) | 12.5 µl | -80°C |
| 5x PARPtrap™ assay buffer 2 | 2 x 1 ml | -80°C | |
| 10x NAD+ | 500 µl | -80°C | |
| 79685 | Black 96-well plate | 1 | Room Temp |
384 Reactions
| Catalog # | Name | Amount | Storage |
| 80502 | PARP2, GST-tag* | 2 x 8 µg | -80°C |
| 78297 | Fluorescent labeled nicked oligonucleotide duplex (100 nM) | 2 x 12.5 µl | -80°C |
| 5x PARPtrap™ assay buffer 2 | 4 x 1 ml | -80°C | |
| 10x NAD+ | 2 x 500 µl | -80°C | |
| 79961 | Black 384-well plate | 1 | Room Temp |
*The initial concentration of enzyme is lot-specific and will be indicated on the tube containing the protein.
Note: Kit updated on 08/02/2022. The revised kit now contains #78297 at concentration 100 nM and requires an added dilution step. Please verify the concentration of #78297 and make sure the corresponding datasheet is being used.
1. Murai, J. et al. Molecular Cancer Therapeutics 2014. 13: 433-443
2. Murai, J. al. Cancer Research 2012. 72: 5588-5599
3. Zandarashvili, L. et al. Science 2020. 368(6486): 30-31
Powered by Bioz
San Diego, CA 92121
858-202-1401
Products are for Research Use Only
COMPANY
Copyright © 2024
BPS Bioscience, Inc.
All rights reserved.
Diagrams on this website are created with BioRender.com.