Proteasome-Glo Caspase-Like Assay
The Proteasome-Glo Cell-Based Assays are 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. The Proteasome-Glo 3-Substrate System consists of three homogeneous bioluminescent assays in an enzyme-based format (each of these three assays is also 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).