The Caspase-3 colorimetric protease assay includes substrate and optimized buffers. The substrate, DEVD-pNA, is composed of the chromophore p-nitroanilide (pNA), and a synthetic tetrapeptide, DEVD (Asp-Glue-Val-Asp), which is the upstream amino acid sequence of the Caspase-3 cleavage site in PARP. Upon cleavage of the substrate by Caspase-3 or related caspases, free pNA light absorbance can be quantified using a spectrophotometer or a microplate reader at 400 or 405 nm. Comparison of the absorbance of pNA from apoptotic sample with an uninduced control allows determination of the fold increase in Caspase-3 activity.
Includes 96-well plate, cuvettes
Apoptosis, Apoptosis Related Factors, Caspase Activation, Cell Analysis, Cell Viability, Proliferation and Function, Enzyme and Protein Activity Assays, Protease/Peptidase Activity, Proteins, Expression, Isolation and Analysis
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