The Cell-Based Proteasome-Glo™ Assays are homogeneous, luminescent assays that individually measure the chymotrypsin-like, trypsin-like and caspase-like protease activities associated with the proteasome complex in cultured cells. These assays can also be used to screen inhibitor libraries and monitor proteasome-regulated protein degradation in cells.

• Simple 'add-mix-measure' protocol
• Chymotrypsin-like, trypsin-like and caspase-like activity data directly from cellular environment
• Results in 10 to 30 minutes after reagent addition
• Luminescent assay avoids inherent fluorescent background signals, providing excellent signal-to-background ratio
• Can be miniaturised to 384-well format

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 optimised for its specific proteasome activity and luciferase activity. The reagents are added to cells in culture, and the proteasome cleaves the substrates, releasing luciferin, which is consumed by luciferase, producing 'glow-type' luminescence correlating to enzyme activity or inhibition. The 26S proteasome is a 2,5 MDa multiprotein complex found in all eukaryotic cells.

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). The simplified method minimises handling steps and makes the assays amenable to automation.