Determination of Significance:
Caspase is a family of proteases involved in the process of apoptosis, including more than 10 members. Caspase-3 is the most important terminal protease in the process of apoptosis, and it is also the most studied caspase; it activates pro-caspase-2,6,7,9, specifically hydrolyzes a variety of key apoptotic proteins, such as PARP, and mediates chromatin condensation, apoptotic body formation and nuclear DNA fragmentation.
Measurement Principle:
The caspase-3 colorimetric assay is based on the hydrolysis of the peptide substrate DEVD-pNA (Asp-Glu-Val-Asp-p-nitroanilide) by caspase-3, resulting in the release of the p-nitroaniline (pNA) moiety. p-Nitroaniline has a high absorbance at 405 nm. The activity of Caspase can be calculated by detecting pNA. This kit is suitable for mammalian tissue and cell.
Self Provided:
Spectrophotometer/Microplate reader, 100μL cuvette/ 96-well plate, centrifuge, water bath / incubator, adjustable pipette, mortar / homogenizer, ice and distilled water
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