USP15 Inhibitor Screening Assay Kit

The USP15 Inhibitor Screening Assay Kit is a 96-well format fluorogenic assay kit designed to measure the activity of the deubiquitinating (DUB) enzyme USP15 (ubiquitin specific peptidase 15) for screening and profiling applications. The kit contains enough purified USP15, Ubiquitinated-AMC substrate, and assay buffer for 100 reactions. To determine the effect of an inhibitor on USP15 activity, the enzyme should be preincubated with or without the test inhibitor prior to adding the Ub-AMC substrate to the reaction. The assay was functionally validated using Ub-Aldehyde, a potent inhibitor of the DUB subfamilies Ubiquitin C-terminal Hydrolases (UCHs), Ubiquitin-Specific Proteases (USPs), Ovarian Tumor Proteases (OTU), and Machado-Josephin Domain (MJD) proteases. Ubiquitin-AMC is a fluorogenic substrate for ubiquitin hydrolases, with a C-terminus derivatization of ubiquitin with 7-amido-4-methylcoumarin (AMC). In the conjugated form, the energy emitted from fluorochrome AMC is quenched. Upon proteolysis, AMC is no longer quenched and emits fluorescence with an lambdaexcitation/lambdaemission maxima of 350/460 nm. The increase in fluorescence is proportional to the DUB activity. Need us to run inhibitor screens or profile your compounds against USP15? Check out our Protease Screening Services or Ubiquitination Screening Services.