Determination of Significance:
Caspase is a family of proteases involved in the process of cell apoptosis, including more than 10 members. Caspase-9, also known as ICE-LAP6 or Mch6, can form a complex with cytochrome c and Apaf1 and to be activated, further activate caspase-3, the most critical enzyme for cell apoptosis, thereby triggering the apoptosis cascade. Caspase-9 is an important upstream caspase in the process of apoptosis signal transduction, and its activation can be regulated by phosphorylation.
Measurement Principle:
The caspase-9 colorimetric assay is based on the hydrolysis of the peptide substrate acetyl- Leu-Glu-His-Asp-p-nitroanilide (LEHD-pNA) by caspase-9, 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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