Caspase-Glo® 9 Assay Systems

The Caspase-Glo® 9 Assay is a homogeneous luminescent assay that measures caspase-9 activity. The assay provides a proluminogenic caspase-9 substrate in a buffer system optimized for caspase activity, luciferase activity and cell lysis. The addition of a single Caspase-Glo® 9 Reagent in an "add-mix-read" format results in cell lysis, followed by caspase cleavage of the substrate and generation of a "glow-type" luminescent signal. The signal generated is proportional to the amount of caspase activity present. The Caspase-Glo® Reagent relies on the properties of a proprietary thermostable luciferase (Ultra-Glo™ Reco...

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The Caspase-Glo® 9 Assay is a homogeneous luminescent assay that measures caspase-9 activity. The assay provides a proluminogenic caspase-9 substrate in a buffer system optimized for caspase activity, luciferase activity and cell lysis. The addition of a single Caspase-Glo® 9 Reagent in an "add-mix-read" format results in cell lysis, followed by caspase cleavage of the substrate and generation of a "glow-type" luminescent signal. The signal generated is proportional to the amount of caspase activity present. The Caspase-Glo® Reagent relies on the properties of a proprietary thermostable luciferase (Ultra-Glo™ Recombinant Luciferase), which generates the stable "glow-type" luminescent signal and improves performance across a wide range of assay conditions.

The system now includes a separate vial of a protease inhibitor, MG-132 Inhibitor, which may be used to reduce background, thus improving the performance of the Caspase-Glo® 9 Assay in cell-based applications.

Features - Benefits

• Simplify Apoptosis or Caspase Detection: The homogeneous "add-mix-read" protocol makes the assay easy to automate; simply add an equal volume of reagent to sample volume.
• Use Less Enzyme: 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 fluorescent-based assays. Maximum sensitivity is achieved in as little as 0.5–1 hour.
• Rely on a Performance-Tested Assay: The assay delivers excellent Z´ factors in cell and purified enzyme models.
• Get More Information: Multiplex with other cell-based assays from Promega.
• Experience Improved Caspase-9 Selectivity: The Caspase-Glo® 9 Assay uses a luminogenic substrate containing the LEHD sequence, which has been shown to be selective for caspase-9. The assay includes an optional proteasome inhibitor (MG-132), which when added to the Caspase-Glo® 9 Reagent significantly reduces nonspecific background in cell-based assays.
• 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/myway/

Applications

• Caspase-9 activity determination.
• Caspase-9 inhibition determination (IC₅₀).
• Caspase-9 activation determination (EC₅₀).

For more information, see the Protocols & Applications Guide.

Notes

Using 100μl of Caspase-Glo® Reagent per assay in a 96-well format, Cat.# G8210 contains sufficient reagents for 25 assays; Cat.# G8211, 100 assays; Cat.# G8212, 1,000 assays, respectively. Using 25μl of Caspase-Glo® Reagent per assay in a 384-well format, Cat.# G8210 contains sufficient reagents for 100 assays; Cat.# G8211, 400 assays; Cat.# G8212, 4,000 assays, respectively.

References

1. Thornberry, N.A., Chapman, K.T. and Nicholson, D.W. (2000) Methods Enzymol. 322, 100–10.
2. Garcio-Calvo, M. et al. (1999) Cell Death Differ. 6, 362–9.

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