| Assay Method Information | |
| | procaspase-3 activation |
| Description: | Procaspase-3 was recombinantly expressed in E. coli with an N-terminal His-6 tag (SEQ ID NO: 29) and purified. Immunoblotting was performed with an anti-His-6 antibody (His-6 disclosed as SEQ ID NO: 29). In the absence of PAC-1 no maturation of procaspase-3 is observed. In the presence of 100 μM PAC-1, cleavage to generate the p19 fragment is observed within 1 h, and >50% cleavage is observed after 4 h. PAC-1 is also effective at 5 μM in this assay. FIG. 2A) Activation of mutants in the safety catch region of procaspase-3 by PAC-1. PAC-1 has an EC50 for activation of 0.22 μM on wild type procaspase-3 (DDD), and corresponding EC50 values of 2.77 μM (DAD), 113 μM (DDA), and 131 μM (ADD) for the mutants. FIG. 2B) PAC-1 activates procaspase-7 with an EC50 of 4.5 μM. FIG. 2C) Dependence of PAC-1 activation of procaspase-3 on pH. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |