Science Catalog 2012 Life Cell Signaling DPPIV-Glo™ Protease Assay Product DPPIV-Glo™ Protease Assay For Laboratory Use. Description: The DPPIV-Glo™ Protease Assay is a homogeneous, luminescent assay that measures dipeptidyl peptidase IV (DPPIV) activity. DPPIV is a serine protease that cleaves N-terminal dipeptides from polypeptides with L-proline or L-alanine at the penultimate position. The DPPIV-Glo™ Assay provides a proluminescent DPPIV substrate, Gly-Pro-aminoluciferin, in a buffer system optimized for DPPIV and luciferase activities. The addition of a single DPPIV-Glo™ Reagent in an “add-mix-measure” format results in DPPIV cleavage of the substrate and generation of a “glow-type” luminescent signal produced by the luciferase reaction. In this homogeneous, coupled-enzyme format, the signal is proportional to the amount of DPPIV activity present. The assay is designed for use with purified enzyme preparations. Features: • Simplified Method: The homogeneous “add-mix-measure” protocol makes the assay highly amenable to automation. • Greater Sensitivity: The assay is more sensitive than fluorescent-based DPPIV assays. In contrast to fluorescent assays, the luminescent assay avoids inherent fluorescent background signals and thus provides excellent signal-to-background readings. The assay is linear over more than 3 logs of DPPIV concentration and can detect less than 1pg/ml enzyme. • Faster Results: The maximum signal (and maximum sensitivity) of the assay is reached in as little as 30 minutes after reagent addition and, unlike fluorescent assays, is not dependent on accumulation of cleaved product. • Amenable to Batch Processing: The stability of the signal allows plates to be read over an extended period of time. Storage Conditions: Store components at –20°C protected from light. Protocol DPPIV-Glo™ Protease Assay Technical Bulletin Part# TB339 Size Cat.# Price (Fr) 10 ml G8350 519.00 50 ml G8351 1817.00 Proteasome-Glo™ Assays Product Proteasome-Glo™ Chymotrypsin-Like Assay Proteasome-Glo™ Trypsin-Like Assay Proteasome-Glo™ Caspase-Like Assay Proteasome-Glo™ 3-Substrate System For Research Use Only. Not for Use in Diagnostic Procedures. Description: The Proteasome-Glo™ Assays are homogeneous, luminescent assays that individually measure the chymotrypsin-like, trypsin-like and caspase-like protease activities associated with the proteasome in an enzyme-based format. The 26S proteasome is a 2.5MDa multiprotein complex found in all eukaryotic cells. Proteasome-Glo™ Cell-Based Assays provide luminogenic proteasome substrates in buffers optimized for cell permeabilization, proteasome activity and luciferase activity. Addition of the Proteasome-Glo™ Cell-Based Reagent in an “add-mix-measure” format results in proteasome cleavage of the substrate and rapid generation of a luminescent signal produced by the luciferase reaction. The three luminogenic substrates used to monitor specific protease activities include: Suc-LLVY-aminoluciferin for chymotrypsin-like, Z-LRR-aminoluciferin for trypsin-like, and Z-nLPnLD-aminoluciferin for caspase-like activity. Each luminogenic substrate is added to a buffer system optimized for its specific proteasome activity and luciferase activity. The reagents are added to test samples containing proteasome enzyme that cleaves the substrates, releasing luciferin, which is consumed by luciferase, producing “glow-type” luminescence correlating to enzyme activity or inhibition. The Proteasome-Glo™ 3-Substrate System consists of three homogeneous bioluminescent assays in an enzyme-based format (each of these three assays also is available separately). The Proteasome-Glo™ Cell-Based 3-Substrate System consists of three homogeneous bioluminescent assays that measure the three proteolytic activities associated with the proteasome in a cell-based format (each of these three assays also is available separately). Features: 1,000 10,000 100,000 100 10 0.1 1 0.0001 0.001 0.01 DPPIV (ng/ml) Sensitivity of the DPPIV-Glo™ Protease Assay compared to a fluorescent assay. 0.1 1 Gly-Pro-aminoluciferin Gly-Pro-AMC • Simplified Method: The “add-mix-measure” protocol minimizes handling steps and makes the assays amenable to automation. • Faster Results: Maximum sensitivity is reached 10–30 minutes after reagent addition. • Greater Sensitivity: The luminescent assay format avoids inherent fluorescent background signals, providing excellent signal-to-background readings. The assays are miniaturizable to 384-well format. Storage Conditions: Store the Proteasome-Glo™ Assay components at –20°C. Protocol Proteasome-Glo™ Assay Systems Technical Bulletin Part# TB349 Size Cat.# Price (Fr) 10 ml G8621 519.00 50 ml G8622 1817.00 10 ml G8631 519.00 50 ml G8632 1817.00 10 ml G8641 519.00 50 ml G8642 1817.00 10 ml G8531 1139.00 50 ml G8532 3987.00 142 For complete and up-to-date product information visit: www.promega.com/catalog Signal-to-Noise Ratio 4799MA Pagina 145
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