FAsformulation Relative caspase-3 activation1 (%) Relative caspase-9 activation1 (%) OA <1 ± 1 <1 ± 1 Cyt c-OA2 48 ± 4 56 ± 5 Cyt c-OA3 26 ± 2 30 ± 3 Cyt c4 100 ± 9 100 ± 3 Cyt c2 73 ± 3 81 ± 4 BSA-OA2 <1 ± 1 <1 ± 2 BSA2 <1 ± 2 <1 ± 1

1The relative activities were calculated from the absorbance changes caused by the caspase-3 and caspase-9 hydrolysis of pNA-substrates, 10 mM DEVD-pNA and 4 mM LEHD-pNA, respectively. pNA formation was monitored colorimetrically at ~ 405 nm. 2These samples were prepared using the original conditions mentioned before in Tris-HCl, pH 8 and 45°C. 3This complex was prepared using the original conditions except by heating at 65°C. 4Native Cyt c. The values in the table are the averages of triplicate measurements and the error values are the calculated S.D.

Table 2: Capability of various preparations to activate caspase-3 and -9 in cell-free assays.