| Solution Information | help | |
| Enzyme: | Caspase-3 | |
| inhibitor: | BDBM50445 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: The purpose of this assay is to determine pro-caspase 3 dose response activation curves for compounds identified as active in a set of experiments entitled, "Absorbance-based confirmation cell-based high throughput confirmation assay for activators of procaspase-3" (PubChem AID 488954), and that were inactive in "Counterscreen for procaspase-3 activators: absorbance-based cell-based high throughput confirmation assay for activators of procaspase-7" (PubChem AID 488941). This assay employs a procaspase 3 mutant enzyme, D9A/D28A/D175A (called PC-3 D3A) which is unable to autoproteolyze itself because its aspartic acid cleavage sites have been mutated to alanines. The mutant has a fully functional active site that can process the peptidic Ac-DEVD-pNA chromogenic substrate. Cleavage of substrate by procaspase 3 hydrolyzes the bond between the aspartic acid and p-nitroaniline, leading to release of yellow p-nitroaniline and an increase in well absorbance at 405 nm. In this | |
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