USP5 Inhibitor Screening Assay Kit

Catalog #

78832

$840 *

Size: 96 reactions

Qty

Bulk/Custom

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*US Pricing only. For international pricing, please contact your local distributor.

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Description

The USP5 Inhibitor Screening Assay Kit is a 96-well format fluorogenic assay designed to measure the activity of the deubiquitinating (DUB) enzyme USP5 for screening and profiling applications. To determine the effect of an inhibitor on USP5 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 based on the 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.

Need us to run inhibitor screens or profile your compounds against USP5? Check out our Protease Screening Services or Ubiquitination Screening Services.

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Synonyms

Deubiquitinating enzyme 5, Isopeptidase T, Ubiquitin thioesterase 5, Ubiquitin-specific-processing protease 5, ISOT, USP-5

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Product Data Gallery

USP5 activity is inhibited by Ub-Aldehyde.

Product Info

Storage and Usage

Citations

Assay Kit Format

Fluorogenic

Supplied As

The kit contains enough purified USP5 protein, Ubiquitinated-AMC substrate, and assay buffer for 100 reactions.

Materials Required But Not Supplied

Adjustable micropipettor and sterile tips
Fluorescence plate reader

Format

Catalog #	Name	Amount	Storage
80355	USP5, FLAG Tag*	>1 µg	-80°C
81150	Ub-AMC Substrate	5 µl	-80°C
79274	10x PR-01 Assay Buffer	3 x 1 ml	-80°C
	0.5 M DTT	200 µl	-80°C
79685	96-well black microplate	1	Room Temp

* The concentration of protein is lot-specific and will be indicated on the tube containing the protein.

UniProt #

P45974

Background

Ubiquitin specific peptidase 5 (USP5) 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. DUBs are a new therapeutic target for neurodegenerative diseases, cancer, diabetes, and autoimmune pathologies.

Storage/Stability

This assay kit will perform optimally for up to 6 monthsfrom date of receipt when the materials are stored as directed.

Applications

Enzyme kinetics studies and screening small molecule inhibitors for drug discovery and high-throughput screen
applications.

Shipping Temperature

-80°C

Notes

Troubleshooting Guide
Visit assay-kits-faq for detailed troubleshooting instructions. For all further questions, please email [email protected]

Warnings

Avoid freeze/thaw cycles

Disclaimers and Limitations

BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.

End-users should immediately report any issues to [email protected] upon receipt of the kit, such as missing components, damaged vials, no dry ice, etc.

Scientific Category

Ubiquitination

Regulatory Status

Research Use Only