Cell Signaling Caspase-Glo® 2 Assay Systems Product Caspase-Glo® 2 Assay For Laboratory Use. Description: The luminescent Caspase-Glo® 2 Assay measures caspase-2 activity in a homogeneous format. Caspase-2 is a member of the cysteine aspartic acid-specific protease family. The Caspase-Glo® 2 Assay provides a luminogenic substrate (Z-VDVAD-aminoluciferin) in a reagent optimized for caspase-2 and luciferase activity. A single reagent is added to test samples, resulting in caspase cleavage of the substrate and generation of a glow-type luminescent signal produced by luciferase. Luminescence is proportional to the amount of caspase activity present. The assay system may be used with purified enzyme preparations and is ideal for automated high-throughput screening of inhibitors. Features: • Broad Dynamic Range: The assay is linear over four logs of caspase-2 concentration and can detect caspase-2 activity at concentrations as low as 0.2mU/ml. • High-Quality Assay: The assay demonstrates an excellent Z´-factor value of 0.85 in 384-well plates using 0.05U/ml of enzyme. • Increased Accuracy: The superior sensitivity over fluorescence-based caspase assays allows inhibitor studies to be performed below the Km. • Batch Processing Capability: The coupled-enzyme, homogeneous format results in a continuous signal, providing excellent stability and allowing plates to be read over an extended period of time. Luminometers with reagent injectors are not required. Storage Conditions: Store at –20°C. Protocol Caspase-Glo® 2 Assay Technical Bulletin Part# TB365 For Laboratory Use. Description: The homogeneous Caspase-Glo® 3/7 Assay measures caspase-3/7 activities using 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 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 than fluorescent- or colorimetric-based assays. Features: • Simplify Apoptosis or Caspase Detection: The “add-mix-measure” protocol makes the assay easy to automate. • Use Less Sample: Excellent signal-to-noise ratios and superior sensitivity allow 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. Maximum sensitivity 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 time period; no injectors required. • Get More Information: Multiplex with other 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/myway/ Caspase-Glo® Reagent Storage Conditions: Store at –20°C. Protocol Caspase-Glo® 3/7 Assay Technical Bulletin Add equal volume of reagent to samples. Mix. Incubate. Measure Luminometer Schematic diagram of the Caspase-Glo® Assay protocol. 160,000 12,000 10,000 8,000 6,000 4,000 2,000 00 200 400 600 800 luminescence. 140,000 120,000 100,000 80,000 60,000 40,000 20,000 0 Cell # The Caspase-Glo® 3/7 Assay output is linear over a broad range of cell numbers. Jurkat cells were treated with anti-Fas mAb for to induce apoptosis or were left untreated. Anti-Fas-Treated Cells Untreated Cells Part# TB323 Size Cat.# Price (Fr) 10 ml G0940 809.00 50 ml G0941 2832.00 Caspase-Glo® 3/7 Assay Systems Product Caspase-Glo® 3/7 Assay Size Cat.# Price (Fr) 2.5 ml G8090 231.00 10 ml G8091 809.00 10 × 10 ml G8093 Pls. Enq. 100 ml G8092 4045.00 2 For complete and up-to-date product information visit: www.promega.com/catalog 27 Cell Health Assays 4689MB Luminescence (RLU) Background Subtracted 4098MB05_3A 1,000 0 2,000 3,000 4,000 5,000 6,000 7,000 8,000 9,000 10,000 Pagina 30
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