PARP3 Chemiluminescent Assay Kits

The PARP3 Chemiluminescent Assay Kit is designed to measure the activity of PARP3 (poly- (ADP-ribose) polymerase 3) for screening and profiling applications. The PARP3 assay kit comes in a convenient 96-well or 384-well format, with enough recombinant purified PARP3 enzyme, reaction substrates and PARP assay buffer for 100 or 400 enzyme reactions.Figure 1. PARP3 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 PARP3 enzyme and an activated DNA template 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 PARP3 activity.Need us to run inhibitor screens or profile your compounds against PARP3? Check out ourPARP/PARPTrap™ Screening ServicesorDNA Replication and Repair Screening Services.This product has beencited 4 times.

Product Specifications

Background

PARP3, also known as poly- (ADP-ribose) polymerase 3 or NAD+ADP-ribosyltransferase 3, 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. PARP3 is involved in programmed and stress-induced double strand break repair via classical NHEJ (non-homologous end-joining) . In the absence of PARP3, DNA repair is less efficient, and cytotoxicity is potentiated when DNA damaging insults occur. It has been linked to cancer, with upregulation of PARP3 occurring in TGFβ (transforming growth factor β) -induced cell EMT (epithelial to mesenchymal transition) in breast cancer. It also promotes cell stemness. The use of inhibitors towards PARP3 can refrain cells from EMT. In addition, the mTORC2 (mTOR complex 2) signaling pathway is impacted, contributing to synthetic lethality in cells deficient in a complementary DNA repair pathway, such as in BRCA1 (breast cancer type 1 susceptibility protein) - triple negative breast cancers (TNBC) . PARP3 acts in synergy with PARP1 in the response to ionizing radiation, and the use of inhibitors for both PARP1 and PARP3 also provides an opportunity to reduce the concentration of both and decrease the radiation regimen.

UniProt

Q9Y6F1

Applications

Study enzyme kinetics and screen small molecule inhibitors for drug discovery and high throughput screening (HTS) applications.

Format

96 reactionsCatalog #NameAmountStorage80503PARP3, GST-Tag*2 x 1.25 µg-80°C520295x Histone Mixture1 ml-80°C78366PARP Substrate Mixture 14 x 250 µl-80°C8060210x PARP Assay Buffer1 ml-20°C79743Blocking Buffer 325 ml+4°C80605Activated DNA20 µl-80°C0.5 M DTT200 µl-20°C80611Streptavidin-HRP100 µl+4°C79670ELISA ECL Substrate A (translucent bottle) 6 mlRoom TempELISA ECL Substrate B (brown bottle) 6 mlRoom Temp7983796-well module plate1Room Temp*The concentration of the protein is lot-specific and will be indicated on the tube.384 reactionsCatalog #NameAmountStorage80503PARP3, GST-Tag*4.5 µg-80°C520295x Histone Mixture2 x 1 ml-80°C78366PARP Substrate Mixture 18 x 250 µl-80°C8060210x PARP Assay Buffer2 x 1 ml-20°C79743Blocking Buffer 32 x 25 ml+4°C80605Activated DNA2 x 20 µl-80°C0.5 M DTT2 x 200 µl-20°C80611Streptavidin-HRP2 x 100 µl+4°C79670ELISA ECL Substrate A (translucent bottle) 2 x 6 mlRoom TempELISA ECL Substrate B (brown bottle) 2 x 6 mlRoom Temp78188384-well white plate1Room Temp*The concentration of the protein is lot-specific and will be indicated on the tube.

Shipping Conditions

-80°C

Storage Conditions

This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.

Notes

Troubleshooting GuideVisitbpsbioscience.com/assay-kits-faqfor detailed troubleshooting instructions. For further questions, please email[email protected]