The ApoAlert Caspase Assay Kits provide a simple and convenient way to detect caspase protease activity using fluorometric or colorimetric methods. The assay kits detect caspase activation by assaying for the cleavage of a fluorescent or a colorimetric substrate. The entire protocol—from collection of cells to fluorometric or colorimetric results—takes only 90 minutes.
Caspase colorimetric & fluorescent assay kits
The caspase-3 colorimetric assays measure the proteolytic cleavage of the chromophore p-nitroanilide (pNA) by caspase-3. Liberated pNA can be monitored colorimetrically by absorbance at 405 nm.
The caspase-9/6 fluorescent assay detects the shift in fluorescence of 7-amino-4-methoxy coumarin (AMC). Upon cleavage, AMC fluorescence can be measured using a 380-nm excitation filter and a 460-nm emission filter. Fluorescence detection is highly sensitive and can be used to measure even very small amounts of active caspase.
The caspase-3 fluorescent assays detect the shift in fluorescence emission of 7-amino-4-trifluoromethyl coumarin (AFC). After the substrate is cleaved by the protease, AFC emits a yellow-green fluorescence at 505 nm if excited at 400 nm.
Overview
- Measure one of the earliest indicators of apoptosis—caspase proteolytic activity
- Perform the assay directly on cell lysates
- Fast and easy—caspase colorimetric assays and fluorescent assays
- Easily formatted for high-throughput applications
- Works with as few as 1 x 106 adherent cells or cells in suspension
- No substrate handling
Applications
- Measure caspase activity in cells
- Monitor apoptotic events at a cell population level
