PARP12 Chemiluminescent Assay Kit
The PARP12 Chemiluminescent Assay Kit is designed to measure the activity of PARP12 (poly-(ADP-ribose) polymerase 12) for screening and profiling applications. The PARP12 assay kit comes in a convenient 96-well format, with enough recombinant purified PARP12 enzyme (amino acids 500-700), reaction substrates, and PARP assay buffer for 100 enzyme reactions.
Figure 1. PARP12 Chemiluminescent Assay Kit schematic.
Histone proteins are coated on a 96-well plate. Next, a biotinylated NAD+ mix (termed PARP Substrate Mixture) is incubated with the PARP12 enzyme in an optimized assay buffer. The plate is then treated with streptavidin-HRP followed by addition of the ELISA ECL substrate to produce chemiluminescence that can be measured using a chemiluminescence reader. The chemiluminescence signal is proportional to the PARP12 activity.
Need us to run inhibitor screens or profile your compounds against PARP12? Check out our PARP/PARPTrap™ Screening Services or DNA Replication and Repair Screening Services.
- 1x PBS (Phosphate Buffer Saline) Buffer
- PBST Buffer (1x PBS, containing 0.05% Tween-20)
- Luminometer or microplate reader capable of reading chemiluminescence
- Adjustable micropipettor and sterile tips
- Rotating or rocker platform
| Catalog # | Name | Amount | Storage |
| 80513 | PARP12, His-GST-Tag* | 27 µg | -80°C |
| 52029 | 5x Histone Mixture | 1 ml | -80°C |
| 78371 | PARP Substrate Mixture 2 | 4 x 250 µl | -80°C |
| 80602 | 10x PARP Assay Buffer | 1 ml | -20°C |
| 79743 | Blocking Buffer 3 | 25 ml | +4°C |
| 0.5 M DTT | 200 µl | -20°C | |
| 80611 | Streptavidin-HRP | 100 µl | +4°C |
| 79670 | ELISA ECL Substrate A (translucent bottle) | 6 ml | Room Temp |
| ELISA ECL Substrate B (brown bottle) | 6 ml | Room Temp | |
| 79837 | 96-well module plate | 1 | Room Temp |
*The concentration of the protein is lot-specific and will be indicated on the tube.
PARP12, also known as poly-(ADP-ribose) polymerase 12 or ARTD12 (ADP-Ribosyltransferase Diphtheria Toxin-Like 12), is part of the PARP family. ADP ribosylation, which is the addition of an ADP-ribose to a protein, is a reversible post-translational modification of proteins mostly involved in the DNA Damage Response (DDR) pathway. Mono-ADP-ribosylation (termed MARylation) is the addition of a unit of ADP-ribose. PARP12 is involved in regulation of cell growth and survival, maintenance of cell polarity in epithelial cells and anti-viral responses. PARP12 is involved in the pathways that lead to viral degradation in cells but can also enhance NF-κB activation and inflammatory responses. Abnormal activity of PARP12 has been linked to cardiovascular and inflammatory diseases and cancer. Studies in HCC (hepatocellular carcinoma) cells indicated that PARP12 can act as a tumor suppressor and be involved in EMT (epithelial-mesenchymal transition). A deeper understanding of the role of PARP12 in health and disease will increase our knowledge on the PARP family of proteins, and lead to the development of more effective therapies for PARP-related diseases.
Kerr C., et al., 2023 bioRxiv: 2023.06.16.545351.
Grimaldi G., et al., 2022 Proc Natl Acad Sci USA 119(1): e2026494119.
Shao C., et al., 2018 Cell Death & Disease 9: 856.
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