Light-based detection method provides the superior sensitivity and speed, while avoiding the variability of fluorescent-based assays associated with autofluorescence and fluorescent interference
The new Caspase-Glo 3/7 assay combines outstanding sensitivity with unprecedented speed and simplicity in apoptosis detection. The kit complements the existing range of apoptosis, cell viability and cytotoxicity assays available from Promega and combines caspase activity with bioluminescence-based detection. It is this light-based detection method that provides the superior sensitivity and speed, while avoiding the variability inherent in fluorescent-based assays associated with autofluorescence and fluorescent interference.
The kit can be used to measure caspase 3/7 activity in apoptotic cells or to screen for caspase inhibitors in purified enzyme assays.
With results in as little as 30 minutes and sensitivity of less than 50 apoptotic cells per well, this simple 'add and read' format enables laboratories to precisely monitor toxic effects of compounds at high throughput capacity or one sample at a time. The assay provides a pro-luminescent caspase-3/7 DEVD aminoluciferin substrate in a reagent selectively optimised for caspase-3/7 activity.
Addition of this single reagent results in lysis followed by caspase cleavage of the DEVD.
This in turn generates a 'glow type' luminescent signal in the presence of luciferase.
The signal is proportional to caspase activity.
This luminescent assay provides excellent signal to noise ratios due to the small amounts of background luminescence, allowing cost reductions through use of fewer cultured cells or less purified caspase if screening for caspase inhibitors. Additionally, the kit is applicable for high-throughput use and automation.
A stable luminescent signal allows for batch processing of plates in 96 or 384 well format, no injectors required.
Protocols are available for the Biomek 2000 and Biomek FX.
