Determination of Significance:
Caspase is a family of proteases involved in the process of cell apoptosis, including more than 10 members. Caspase-8, also known as FLICE、MACH or Mch5. It is usually in the form of zymogen and activated during apoptosis. It is considered to be the upstream caspase of apoptosis transduction. During Fas receptor and TNFR-1 mediated apoptosis, Caspase-8 is activated to form dimer, and then downstream caspase-4, 6, 9 and 10 are activated.
Measurement Principle:
The caspase-8 colorimetric assay is based on the hydrolysis of the peptide substrate Ac-IETD-pNA (N-acetyl-Ile-Glu-Thr-Asp-p-nitroanilide) by caspase-8, 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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