This ELISA based assay detects poly (ADP-ribose) deposited by PARP-1 onto immobilized histones in a 96-well format. An anti-PAR monoclonal detecting antibody followed by addition of a goat anti-rabbit IgG-HRP secondary and a chemiluminescent HRP substrate yields relative light units (RLU) that correlates with PARP-1 activity. During apoptosis PARP-1 is cleaved with a resulting drop in activity. Etoposide is included as a control apoptosis inducer.
- 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 hrs
- Measure PARP-1 activity in cell extracts before and after apoptosis.
- For the screening of candidate PARP-1 inhibitors and determination of IC50 values.