TheProteasome-Glo™ Assaysare homogeneous, luminescent assays that individually measure the chymotrypsin-like, trypsin-like and caspase-like protease activities associated with the proteasome in a purified enzyme-based format. The 26S proteasome is a 2.5MDa multiprotein complex found in all eukaryotic cells. Adding 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.
TheProteasome-Glo™ 3-Substrate Systemconsists 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 Systemconsists 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 - Benefits
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.
Applications
Measuring the chymotrypsin-like, trypsin-like and caspase-like activities of the proteasome in enzyme preparations.
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