| Assay Method Information | |
| | Caspase Catalytic Activity Assay |
| Description: | Experiments were performed in a 384-well format (Greiner no. 781207) as per the conditions noted here. Caspase-1: 2.5 nM enzyme, 6.5 mM WEHD substrate, ECB; Caspase-3: 200 nM enzyme, 3.3 mM DEVD substrate, SCB; Caspase-4: 1 nM enzyme, 10 mM LEHD substrate, HCB; Caspase-5: 20 nM enzyme, 10 mM LEHD substrate, HCB; Caspase-9: 200 nM enzyme, 6.5 mM LEHD substrate, HCB. ECB (Enzo Caspase Buffer): 50 mM HEPES (pH 7.4), 100 mM NaCl, 0.5% Tween 20, 10 mM DTT, and 10% glycerol; SCB (Standard Caspase Buffer): 20 mM PIPES (pH 7.5), 100 mM NaCl, 1 mM EDTA, 10 mM DTT, and 10% sucrose; HCB (High-Citrate Buffer): 50 mM Tris-HCl (pH 7.5), 1 M sodium citrate, 10 mM DTT, and 10% sucrose. Activity was measured as the change in luminescent signal for at least 30 min. |
| Affinity data for this assay | |
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