Description
A Homogenous, Luminescent Assay That Measures Caspase-8 Activity
The assay provides a proluminogenic caspase-8 substrate in a buffer system optimized for caspase activity, luciferase activity and cell lysis. The addition of a single Caspase-Glo® 8 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® 8 Assay in cell-based applications.
Features and Benefits
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.
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.
Delivers excellent Z´ factors in cell and purified enzyme models.
Notes: Using 100μl of Caspase-Glo® Reagent per assay in a 96-well format, Cat.# G8200 provides sufficient reagents to perform 25 assays; Cat.# G8201, 100 assays; Cat.# G8202, 1,000 assays. Using 25μl of Caspase-Glo® Reagent per assay in a 384-well format, Cat.# G8200 provides sufficient reagents to perform 100 assays; Cat.# G8201, 400 assays; Cat.# G8202, 4,000 assays.
Sensitivity of the Caspase-Glo® 8 Assay in apoptotic Jurkat cells with MG-132 Inhibitor added to the Caspase-Glo® 8 Reagent.
