4.4. DEVDase (Caspase-3) Activity Assay

To evaluate DEVDase (Asp-Glu-Val-Asp-ase) activity, cell lysates were prepared after administration of the appropriate treatment. The assays were performed in 96-well plates by incubating 20 μg of cell lysate in 100 μL of reaction buffer (1% NP-40, 20 mM Tris-HCl [pH 7.5], 137 mM NaCl, and 10% glycerol) containing the caspase substrate (Asp-Glu-Val-Asp-chromophore p-nitroanilide; DEVD-pNA) at 5 μM. The lysates were incubated at 37 °C for 2 h. Thereafter, activity was measured at 405 nm using a spectrophotometer.