Homogeneous Caspases Assay, fluorimetric

Specific, quantitative in vitro determination of caspases in microplates.
During apoptosis, the activation of specific proteases, called Caspases (cysteinyl-aspartic-acid-proteases), is one of the initial intracellular biochemical events. The Homogeneous Caspases Assay can be used to detect caspase activity in life-science research applications:

• for sensitive quantification of activated caspases
• for screening of caspase inhibitors
• as an automatable one-step assay for high-throughput screening

Enzyme activity of natural and recombinant caspase is detected by this homogeneous caspases assay. Activated caspases 2, 3, 6, 7, 8, 9, and 10 can be analyzed with different sensitivity.

The Homogeneous Caspases Assay is a fluorimetric one-step assay for the quantitative in vitro determination of caspase activity in microplates. Cells are cultured in microplates and apoptosis is induced, causing an activation of caspases. After an appropriate incubation period, caspases substrate (DEVD-R110), diluted in incubation buffer, is added and incubated for 1 - 2 hours. During this incubation period, the cells are lysed by the Incubation buffer and the caspases substrate is cleaved in proportion to the amount of activated caspases. The fluorescent dye Rhodamine 110 is released and fluorometrically determined at λ_max = 521nm.
The developed fluorochrome is proportional to the concentration of activated caspases, and can be quantified in a calibration curve.
The total assay time is approximately 1.5 hours (excluding cell culturing).

The Homogeneous Caspases Assay is a fluorimetric assay for the quantitative in vitro determination of caspases activity in microplates, which makes it especially useful for high-throughput screening.

For life science research only. Not for use in diagnostic procedures.

1 kit containing 4 components