During apoptosis, PARP-1 which catalyzes the NAD+ -dependent addition of poly (ADP-ribose) (PAR) onto various cytoplasmic and nuclear proteins, is cleaved from about 116 kDa to 85 kDa. Moreover, this enzyme is a therapeutic target for BRCA1and BRCA2 associated breast cancers. The HT PARP/Apoptosis Assay is ideal for measuring the activity of PARP in cell extracts before and during apoptosis. This ELISA semi-quantitatively detects PAR deposited onto immobilized histone proteins in a 96-well format. An anti-PAR monoclonal antibody, goat anti-mouse IgG-HRP conjugate, and HRP substrate are used to generate a signal. Thus, signals correlate with PARP activity. Etoposide is a topoisomerase II inhibitor that stabilizes this enzyme after it cleaves DNA. It is included as a control apoptosis induce.
FEATURES:
- Colorimetric or Chemiluminescent readout
- 96 well format
- Highly sensitive – detects 0.1 mU PARP ~500 cells
- Dynamic range between 0.1 to 10 mU PARP
- Requires 10-100 ng extract for detection
- Assay Time ~3 hr
APPLICATIONS:
- Measure activity in cells, primary or tumor cells.
- Measure activity before and after apoptosis.
- Measure effect of PARP inhibitors using cell extract.
KIT COMPONENTS:
- PARP-HSA and Buffer
- Anti-PAR Monoclonal Antibody
- Activated DNA
- NAD+
- Histone-Coated Strip Wells
- PAR mAb and Diluent
- HRP Conjugate
- Etoposide
- Colorimetric or Chemiluminescent Detection Reagent
Please see out PARP kit selection guide below.
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