Example of an active caspase 3 protein standard curve.
Example of a dilution series of Jurkat Cell extract, prepared from cells treated for 4 hours with 1 µM staurosporine, in Incubation Buffer in the working range of the assay
Demonstration of assay specificity by induction of active caspase 3 with staurosporine treatment. Jurkat cells were treated for 4 hours with 1 µM staurosporine or drug’s vehicle (DMSO) and extracts at 1000 µg/mL were analyzed using this kit. Active caspase 3 concentrations were interpolated from standard curve.
Example of staurosporine IC50 determination. Lysates corresponding to 2x106 cells/mL or 1x106cells/mL were prepared by direct in-well lysis (without media removal) from Jurkat cells treated with variable doses of staurosporine for 6 hours in a 96-well plate. 100 µL of the lysates were analyzed. Background-subtracted signals are shown in left panel. Active caspase 3 concentrations interpolated from standard curve are shown in right panel. IC50 determined from background-subtracted signals were 0.9 µM and 1.0 µM using, respectively, extracts of 2x106 cells/mL and 1x106 cells/mL.