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. Trevigen's HT PARP/Apoptosis Assay is ideal for measuring the activity of PARP in cell extracts before and during apoptosis. The HT PARP/Apoptosis Assay is an ELISA which 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 colorimetric signal. Thus, absorbance correlates with PARP activity. Etoposide is a topoisomerase II inhibitor that stabilizes this enzyme after it cleaves DNA. It is included as a control apoptosis inducer.
Features:
Chemiluminescent, non-radioactive format
High throughput 96 test size
Sensitivity down to 0.1 mUnits of PARP
Applications: Measuring PARP activity in cell extracts before and during apoptosis
