ApoAlert Caspase Assay Kits
ApoAlert Caspase Assay Kits
Brand: Takara Bio.
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ApoAlert Caspase Assay Kits
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
Detection of caspase activity during apoptosis
Detection of caspase activity during apoptosis. The caspase colorimetric assays and caspase fluorescent assays are based on the same principle—they detect the chromophore or fluorophore that is cleaved from a substrate by the caspase protease.
Fluorometric detection of caspase activity during apoptosis
Fluorometric detection of caspase activity during apoptosis. After protease activation, the activated caspases recognize their respective substrates which are covalently linked to the fluorogenic dye, 7-amino-4-methyl coumarin (AMC). Upon cleavage by the respective caspase, the free dye can be detected using a plate reader with a 380-nm excitation and 460-nm emission filter.
ApoAlert Caspase Profiling Plate layout
ApoAlert Caspase Profiling Plate layout. Caspase profiling assay plates contain immobilized substrate for each of caspases 3, 8, 9, and 2, as shown.
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Detection of caspase activity during apoptosis
Detection of caspase activity during apoptosis. The caspase colorimetric assays and caspase fluorescent assays are based on the same principle—they detect the chromophore or fluorophore that is cleaved from a substrate by the caspase protease.
Fluorometric detection of caspase activity during apoptosis
Fluorometric detection of caspase activity during apoptosis. After protease activation, the activated caspases recognize their respective substrates which are covalently linked to the fluorogenic dye, 7-amino-4-methyl coumarin (AMC). Upon cleavage by the respective caspase, the free dye can be detected using a plate reader with a 380-nm excitation and 460-nm emission filter.
ApoAlert Caspase Profiling Plate layout
ApoAlert Caspase Profiling Plate layout. Caspase profiling assay plates contain immobilized substrate for each of caspases 3, 8, 9, and 2, as shown.
