Ubiquitin, SUMO, ISG15 & NEDD8 (Patented Technology) The Ub-CHOP2-Reporter Deubiquitination Assay consists of ubiquitin fused to a reporter enzyme, as well as a separate reagent substrate for the reporter enzyme. When fused to ubiquitin, the reporter is rendered catalytically inactive. Following cleavage of the Ub-reporter system by the isopeptidase, the free (and now active) reporter subsequently acts upon its substrate. Thus, in this coupled assay, the signal generated by cleavage of the reporter enzyme’s substrate is a quantitative measure of isopeptidase activity. This assay utilizes the two classes of ubiquitin: ubiquitin-like proteins (UBL) and ubiquitin-domain proteins (UDP). The UBLs function as modifiers in a manner analogous to that of ubiquitin. Examples consist of (and are available for this assay) SUMO, Nedd8 (aka Rub1), ISH15, Apg8, APg12, and Fat10. The ubiquitin-domain proteins consist of parkin, RAD23, and DSK2. Service Highlights * Superior to Ub-AMC and FRET-based assays * Rapid and robust readout for DUB activity within 45 minutes with non-radioactive reporter substrates * Amenable to high-throughput screening (HTS) and miniaturization * Assay tests deconjugation of ubiquitin/UBL from a more physiologically relevant protein * Unlike Ub-AMC, the CHOP2-Reporter system does not require excitation in the UV range (reducing the incidence of false-positives)
