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Application: Enzyme kinetics studies and screening small molecule inhibitors for drug discovery and high-throughput screening (HTS) applications.
Background: USP11 (ubiquitin specific protease 11), or ubiquitin carboxyl-terminal hydrolase 11, belongs to the USP family of proteins. USP11 is a DUB (deubiquitinating enzymes) and is involved in removing K-48 and K-63 ubiquitin chains from several substrates. It participates in DNA repair and inflammation, can regulate immune cell differentiation via the NF-κB and TNF (tumor necrosis factor) pathways. It has been linked to the development and progression of several cancer types, such as breast, hepatocellular and ovarian cancer. The development of USP11 specific inhibitors will bring therapeutic benefits in cancer therapy.
Description: The USP11 Inhibitor Screening Assay Kit is a 96-well format fluorogenic assay designed to measure the activity of the deubiquitinating (DUB) enzyme USP11 (ubiquitin specific peptidase 11) for screening and profiling applications. The kit contains enough purified USP11 protein, Ubiquitinated-AMC substrate, and assay buffer for 100 reactions.To determine the effect of an inhibitor on USP11 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 DUB subfamilies ubiquitin C-terminal hydrolases (UCHs), ubiquitin-specific proteases (USPs), Ovarian Tumor Proteases (OTU), and Machado-Josephin domain proteases (MJD).Figure 1: Illustration of the assay principle. Ubiquitin-AMC is a fluorogenic substrate for ubiquitin hydrolases based on the C-terminus derivatization of ubiquitin with 7-amido-4-methylcoumarin (AMC). In conjugated form, the energy emitted from fluorochrome AMC is quenched. Upon proteolysis, AMC is no longer quenched and emits fluorescence with excitation/emission maxima of 350/460 nm. The increase in fluorescence is proportional to DUB activity.Need us to run inhibitor screens or profile your compounds against USP11? Check out our Protease Screening Services or Ubiquitination Screening Services.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Target: USP11
Uniprot: P51784
Warnings: Avoid freeze/thaw cycles. Protect Ub-AMC from direct exposure to light.
Biosafety Level: Not applicable (BSL-1)
References: Liao Y., et al., 2022 Cell Death Discovery 8:292.
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