Caspase glo luminescent based assays

Caspase-3/7activity was analyzed using the Caspase-Glo luminescent-based assays (Promega) according to the manufacturer’s instructions. Briefly, cell were treated with triptolide at the times and concentrations indicated and appropriate Caspase-Glo reagent added to each well. Luminiscence was measured 45 mins after substrate addition. Caspase activity detected was normalized to the number of live cells present detected using the Dojindo cell viability kit.

Caspase-3, 8, 9 activity was analyzed using the Caspase-Glo luminescent-based assays (Promega) according to the manufacturer’s instructions. Cells (3× 10³) were seeded into 96-well white opaque plates and a corresponding optically clear 96-well plate. After 48 hours, cells were treated at concentrations and times indicated. At the end of the incubation, 100 μl of Caspase-Glo reagent was added to each well containing 100 μl of cells in culture medium. Plates were gently mixed and incubated for 45 minutes at room temperature, and luminescence was then read on a luminometer. The corresponding 96-well clear plates were used to measure the number of viable cells with the CCK-8 reagent, and caspase activity was normalized to these values.