DUB-Glo™ Protease Assay (DUB/SENP/NEDP) Technical Manual

Literature # TM319

The DUB-Glo™ Protease Assay (SUB/SENP/NEDP) is a homogeneous, bioluminescent assay that measures the activity of numerous deconjugating enzymes including deubiquitinating (DUB), deSUMOylating (SENP) and deneddylating (NEDP) proteases. These proteases reverse the protein modification by ubiquitin and ubiquitin-like proteins (Ubl proteins) and thus are integral components in the complex mechanisms of posttranslational protein regulation in eukaryotes.
The DUB-Glo™ Protease Assay provides a luminescent substrate, Z-RLRGG-aminoluciferin, in a reagent optimized for protease and luciferase activity. The RLRGG sequence is the C-terminal pentapeptide of ubiquitin. A single DUB-Glo™ Reagent is added to test protease samples, resulting in cleavage of the substrate and generation of a glow-type luminescent signal produced by luciferase. In this coupled-enzyme format, luminescence is proportional to the amount of DUB, SENP or NEDP1 activity present.