Solution Information | help | |
Enzyme: | Caspase-7 | |
inhibitor: | BDBM79436 | |
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 procaspase-7 dose response curves for compounds identified as possible dual procaspase-3/procaspase-7 activators. This biochemical assay employs a procaspase 7 triple-mutant enzyme (D23A/D198A/D206A) (PC-7 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 this substrate by procaspase 7 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 assay, the enzyme is pre-incubated with test compounds, followed by addition of substrate and measurement of well epi-absorbance. As designed, compounds that activate procaspase-7 activity will increase substrate hydrolysis, leading to an increase in well absorbance. Compounds are tested in tripli | |
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