Description
Principle of the Assay: Caspase (Cysteine-requiring Aspartate Protease) is a protease family that plays an important role in the process of apoptosis. Caspase 3 (also known as CPP32, Yama, or apopain) is a member of the CED-3 subfamily of Caspases and is one of the critical enzymes of apoptosis. Caspase 3 is the most studied Caspase in mammalian cells. Caspase 3 can be used to cut proCaspase2, 6, 7, 9, and cut specifically many substrates of Caspase directly, including PPAR (poly ADP-ribose), the inhibitor of Caspase-activated deoxyribonuclease (ICAD), gelsolin and fodrin. Protein shearing mediated by Caspase 3 are important parts of the molecular mechanism of apoptosis. In addition, Caspase 3 also plays a key role in the process of nuclear apoptosis including chromatin condensation and DNA fragmentation as well as cell blebbing. Caspase 3 exists in the form of prozyme in the normal state and has no activity. However, during the apoptosis stage, activated Caspase 3 consists by two large subunits and two small subunits, which cleavages the corresponding substrate of endochylema or cytoplasmic nuclear and eventually leads to apoptosis. This kit is used to conjugate Caspase 3 sequence-specific peptides acetyl-Asp-Glu-Val-Asp p-nitroanilide (AcDEVD-pNA) to yellow group p-nitroaniline (pNA). When the substrate is cut by Caspase 3, the yellow group pNA is dissociated. pNA has an absorption peak at 405nm. Measure the OD value at 405 nm and then Caspase 3 activity can be calculated accordingly: