Caspase 3/7 activity is a cell based assay that measures apoptosis. Commercially available reagents enable high-throughput screening of apoptosis inducing test compounds or contract research organizations can readily perform the compound screening. A brief overview of the protocol is as follows:
- Cells in logarithmic growth are seeded in a 96-well plate
- Depending on the scope of the experiment, the test compound is added either as a single concentration or in a dilution series
- Cells are harvested at various time-points after addition of the test compound (i.e. 6, 12, 24 and 48 hours)
- In the appropriate well for each time-point, the caspase reagent is added to the cells, resulting in cell lysis and caspase cleavage of the fluorescent substrate
- The resultant fluorescent signal is measured using a plate reader (i.e. Molecular Device Spectramax Gemini XPS plate reader)
- The fluorescent signal is proportional to the amount of caspase activity present as a result of treatment by the test article
Altogen Labs, a contract research organization, perform the Caspase 3/7 apoptosis cell-based assay. See here for more information.