USP9X Inhibitor Screening Assay Kit

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Application: Enzyme kinetics and screening small molecule inhibitors for drug discovery and High-Throughput Screening (HTS) applications.

Background: USP9X (Ubiquitin specific peptidase 9 X-linked) belongs to a large group of ubiquitin-specific proteases capable of cleaving ubiquitin from other proteins. These enzymes are also referred to as deubiquitinating peptidases, deubiquitinases (DUBs), ubiquitin proteases, ubiquitin hydrolases or ubiquitin isopeptidases. They contribute to the ubiquitin signaling pathway by countering the signal induced by ubiquitin conjugating enzymes and ligases. USP9X has been linked to atherosclerosis, cancer and disorders of the nervous system. It is involved in multiple crucial cellular functions, such as apoptosis, EMT (epithelial-mesenchymal transition), development of drug resistance, amongst others. It has as targets proteins like MCL-1 (myeloid leukemia cell differentiation 1), SMAD4 (SMAD family member 4) and XIAP (X-linked inhibitor of apoptosis protein). In the context of cancer its role may be context dependent. The development of small molecule inhibitors may thus be a promising therapy for cancers where USP9X acts as oncogene.

Contraindications: The final concentration of DMSO in the assay should not exceed 1%.Compounds that are fluorescent may interfere with the results, depending on their spectral excitation and emission properties.It is recommended that the compound alone is tested to determine any potential interference of the compound with the assay results.

Description: The USP9X Inhibitor Screening Assay Kit is a fluorogenic assay kit designed to measure the activity of the deubiquitinating (DUB) enzyme USP9X (ubiquitin specific peptidase 9 X-linked) for screenin82726-1g and profiling applications. The kit contains enough purified USP9X, Ubiquitinated-AMC substrate, and assay buffer for 96 reactions.To determine the effect of an inhibitor on USP9X 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.Figure 1: Illustration of the assay principle. 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 λexcitation/λemission maxima of 350/460 nm. The increase in fluorescence is proportional to the DUB activity.

Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.

Uniprot: Q93008

Warnings: Avoid freeze/thaw cycles

Biosafety Level: Not applicable (BSL-1)

• Artikelnummer: BPS82726
• Hersteller: BPS Bioscience
• Hersteller Artikelnummer: 82726
• Verpackungseinheit: 96 reactions
• Mengeneinheit: PAK