The Caspase-Glo® 3/7 Assay is a luminescent assay that measures caspase-3 and -7 activities in purified enzyme preparations or cultures of adherent or suspension cells. The assay provides a proluminescent caspase-3/7 substrate, which contains the tetrapeptide sequence DEVD. This substrate is cleaved to release aminoluciferin, a substrate of luciferase used in the production of light. The Caspase-Glo® 3/7 Reagent is optimized for caspase activity, luciferase activity and cell lysis. Addition of the single Caspase-Glo® 3/7 Reagent in an "add-mix-measure" format results in cell lysis, followed by caspase cleavage of the substrate and generation of a “glow-type” luminescent signal. The Caspase-Glo® 3/7 Assay is designed for use with multiwell plate formats, making it ideal for automated high-throughput screening of caspase activity or apoptosis.
The Caspase-Glo® 3/7 Assay provides a homogeneous luminescent assay that measures caspase-3/7 activities. The assay provides a proluminescent caspase-3/7 DEVD-aminoluciferin substrate and a proprietary thermostable luciferase in a reagent optimized for caspase-3/7 activity, luciferase activity and cell lysis. Adding the single Caspase-Glo® 3/7 Reagent in an "add-mix-measure" format results in cell lysis, followed by caspase cleavage of the substrate. This liberates free aminoluciferin, which is consumed by the luciferase, generating a "glow-type" luminescent signal. The signal is proportional to caspase-3/7 activity. The stabilized luciferase and proprietary buffer system improve assay performance across a wide range of assay conditions, and the assay is less likely to be affected by compound interference unlike fluorescent- or colorimetric-based assays. The Caspase-Glo® 3/7 Assay is designed for use with multiwell plate formats using either purified enzyme or cells in culture.
Features - Benefits
Simplify Apoptosis or Caspase Detection: The "add-mix-measure" protocol makes the assay easy to automate; simply add an equal volume of reagent to sample volume.
Use Less Enzyme or Fewer Cells: The low background luminescence results in excellent signal-to-noise ratios and superior sensitivity not achieved by other caspase formats, allowing assays to be performed in 96- or 384-well formats.
Decrease Assay Time: No sample preparation or manipulation required, and no extended incubation times are necessary, as with fluorescence-based assays. Maximum sensitivity is achieved in as little as 0.25–1 hour.
Rely on a Performance-Tested Assay: The assay delivers excellent Z´- factor values in cell and purified enzyme models.
Process Plates in Batch Mode: The extended-glow signal allows the plates to be read over a 3-hour period of time for batch processing; no injectors required.
Get More Information: Multiplex with other cell-based assays from Promega.
Automate This Assay: Validated automated methods available at: www.promega.com/automethods/
Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/custom/
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