Caspase-3 assay Kit for drug discovery

• Cat.No.: Kit-0161
• Product Overview: The CASPASE-3 Assay Kit for Drug Discovery is a complete assay system designed to screen caspase-3 inhibitors. It contains both a colorimetric substrate (DEVD-pNA) and a fluorogenic substrate (DEVD-AMC). Cleavage of the p-nitroanilide (pNA) from the colorimetric substrate increases absorption at 405nm. The fluorescent assay is based on the cleavage of 7-amino-4-methylcoumarin (AMC) dye from the C-terminus of the peptide substrate. Cleavage of the dye from the substrate increases its fluorescence intensity at 460 nm. The assays are performed in a convenient 96-well microplate format. The kit is useful to screen inhibitors of caspase-3, a potential therapeutic target. An inhibitor, DEVD-CHO (aldehyde), is also included as a prototypic control inhibitor. The DEVD amino acid sequence is derived from the caspase-3 cleavage site in PARP.

Specification

• Size: 96 wells
• Storage: Store all components except the microplate at -70°C for the highest stability. The caspase-3 enzyme component must be handled particularly carefully in order to retain maximal enzymatic activity. Defrost it quickly in a RT water bath or by rubbing between fingers, then immediately store on an ice bath. The remaining unused enzyme should be quickly refrozen by placing at -70°C. The enzyme is stable to freeze/thaw cycles x 4. To minimize the number of freeze/thaw cycles, aliquot the caspase-3 into separate tubes and store at -70°C.