PARP2 Homogenous Assay Kit
The PARP2 Homogeneous Assay Kit is designed to measure PARP2 (poly-(ADP-ribose) polymerase 2) activity for screening and profiling applications. The PARP2 Homogeneous Assay Kit comes in a convenient 384-well AlphaLISA® format, with enough purified recombinant PARP2 (amino acids 2-583), biotinylated histone substrate, ADP-Ribose Binding Reagent 1 and assay buffer for 400 enzyme reactions.
Figure 1: PARP2 Homogenous Assay Kit schematic.
A sample containing PARP2 is incubated with a biotinylated histone substrate and NAD+ for one hour. This is followed by the addition of acceptor beads and ADP-Ribose Binding Reagent 1, and finally donor beads. Alpha-counts are then measured. Alpha-counts are directly proportional to PARP2 activity
Need us to run inhibitor screens or profile your compounds against PARP2? Check out our PARP/PARPTrap™ Screening Services or DNA Replication and Repair Screening Services.
- AlphaLISA® Anti-Rabbit IgG Acceptor Beads (Perkin Elmer #AL104C)
- AlphaScreen® Streptavidin-Conjugated Donor Beads (Perkin Elmer #6760002S)
- Optiplate - 384 (Perkin Elmer #6007290)
- AlphaScreen microplate reader
Catalog # | Name | Amount | Storage |
80502 | PARP2, GST-Tag* | 2 µg | -80°C |
5X PP-01 Assay Buffer | 2 x 1 ml | -80°C | |
Biotinylated Histone Substrate | 500 reactions | -80°C | |
78311 | ADP-Ribose Binding Reagent 1 | 10 µl | -80°C |
750 µM NAD+ | 400 µl | -80°C | |
0.5 mM DTT | 200 µl | -20°C | |
4X Detection Buffer 1NP | 2 ml | -20°C |
*The initial concentration of enzyme is lot-specific and will be indicated on the tube containing the protein.
PARP2 (poly(ADP-ribose polymerase 2) is part of the PARP family of proteins, involved in poly(ADP-ribosyl)ation reactions. PARP proteins, mainly PARP1 and PARP2, respond to cellular stress and are involved in DNA repair and transcriptional regulation. PARP2 is linked to oxidative stress, by altering the subcellular distribution of NRF2 (nuclear factor erythroid 2-related factor 2). Under normal conditions NRF2 forms a complex with KEAP1 (Kelch like-ECH-associated protein 1) and it is targeted for degradation by the Cul-3-Rbx1-E3 ligase complex. Under stress conditions KEAP1 releases NRF2, and NRF2 can translocate to the nucleus, a process that requires PARP2. Dysfunction in the cell redox balance and inability to repair DNA contributes to pathologies like inflammation, cancer and cardiovascular diseases. Four PARP1/2 inhibitors are currently used in the clinic for the treatment of BRCA-deficient breast, ovarian, prostate and pancreatic cancer, and the discovery of new inhibitors will lead to new avenues for the treatment of PARP linked diseases.
Gui B., et al., 2019 Proc Natl Acad Sci USA 116 (29): 14573-14582