PARP1 Colorimetric Assay Kit

PARP1, also known as poly- (ADP-ribose) polymerase 1 or NAD+ADP-ribosyltransferase 1, is part of the PARP family, and it is the most abundant member.  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. Poly-ADP-ribosylation (termed PARylation) is the addition of linear or branched chains of ADP-ribose. PARP1 participates in DNA repair by non-homologous end joining (NHEJ), homologous recombination (HR), microhomology-mediated end-joining (MMEJ) and nucleotide excision repair. Dysfunction of DDR pathways can lead to oncogenesis. Overexpression of PARP1 has been found in breast and colon cancer, neuroblastoma, and others. This overexpression can lead to increasing MMEJ, an error-prone DNA repair mechanism, and genome instability leading to cancer. In addition to being involved in DDR, PARP1 is also linked to inflammation and type I diabetes. PARP1 inhibitors have been used in cancer treatment with success. In addition to reducing MMEJ, the use of PARP1 inhibitors can lead to synthetic lethality when homologous recombination repair (HRR) mechanisms are already defective, as in the case of BRCA1 (breast cancer susceptibility protein type 1) and BRCA2 deficient cells. Further understanding of the molecular pathways involving PARP1, and this contribution to disease, will continue to pave the way for new therapies for PARP1-linked diseases.

The PARP1 Colorimetric Assay Kit is designed to measure PARP1 (poly- (ADP-ribose) polymerase 1) activity for screening and profiling applications. The PARP1 assay kit comes in a convenient 96-well format, with enough purified PARP1 enzyme, reaction substrates and PARP assay buffer for 100 enzyme reactions.Figure 1. PARP1 Colorimetric 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 PARP1 enzyme and an activated DNA template in an optimized assay buffer. The plate is then treated with streptavidin-HRP followed by addition of the colorimetric HRP Substrate which generates a colored solution.  The signal generated is proportional to PARP1 activityNeed us to run inhibitor screens or profile your compounds against PARP1? Check out ourPARP/PARPTrap™ Screening ServicesorDNA Replication and Repair Screening Services.This product has beencited 32 times.

P09874

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

Catalog #NameAmountStorage80501PARP1, GST-Tag*1 µg-80°C520295x Histone Mixture1 ml-80°C78366PARP Substrate Mixture 1250 µl-80°C8060210x PARP Assay Buffer1 ml-20°C79743Blocking Buffer 325 ml+4°C80605Activated DNA20 µl-80°C80611Streptavidin-HRP100 µl+4°C79651Colorimetric HRP substrate10 ml+4°C0.5 M DTT200 µl-20°C7996496-well transparent plate1Room Temp*The concentration of the protein is lot-specific and will be indicated on the tube

-80°C

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

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