Caspases, cysteine-aspartic proteases or cysteine-dependent aspartate directed proteases, are a family of cysteine proteases that play essential roles in apoptosis (programmed cell death), necrosis, and inflammation. Caspases are essential in cells for development and most other stages of adult life, and have been termed "executioner" proteins for their roles in cells. Some caspases are also required in the immune system for the maturation of lymphocytes. Failure of apoptosis is one of the main contributions to tumor development and autoimmune diseases. It is also reported that the unwanted apoptosis occurs with ischemia or Alzheimer's disease. The new findings have stimulated interest in caspases as potential therapeutic targets since they were discovered in the mid-1990s.
Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit is designed to monitor cell apoptosis through measuring Caspase 3 activation. Caspase 3 is widely accepted as a reliable indicator for cell apoptosis since the activation of caspase-3 (CPP32/apopain) is important for the initiation of apoptosis. Caspase 3 has substrate selectivity for the peptide sequence Asp-Glu-Val-Asp (DEVD). Z-DEVD-ProRed™ is used in the kit as the fluorogenic indicator for caspase-3 activity. Cleavage of ProRed™ DEVD blocking peptide residue by caspase 3 generates strongly red fluorescent ProRed™ that is fluorimetrically monitored at ~620 nm with excitation at ~530 nm.
Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit provides all the essential components with an optimized assay protocol. The assay is robust, and can be readily adapted for high-throughput assays in a wide variety of fluorescence platforms such as microplate assays. Using 100 µL of reagents per well in a 96-well format, this kit provides sufficient reagents to perform 100 assays. Using 25 µL of reagents per well in a 384-well format, this kit provides sufficient reagents to perform 400 assays.
Key Features of Cell Meter™ Caspase 3/7 Activity Assay Kits: