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. Adding the Proteasome-Glo™ 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.
Homogeneous, luminescent enzyme-based assays (not suitable for cell-based applications)
- Simple "add-mix-measure" protocols
- Results in 10–30 minutes after reagent addition
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 26S proteasome is a 2.5MDa multiprotein complex found in all eukaryotic cells. 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 simplified method minimizes handling steps and makes the assays amenable to automation. The luminescent assay format avoids inherent fluorescent background signals, providing excellent signal-to-background readings. The assays are miniaturizable to 384-well format.