DUB Activity Kit
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Catalog numberSKT-136-96
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Price:
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Size96 well
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Stock availabilityIn Stock
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Kit s descriptionFluorometric detection of DUB activity
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Protein targetDUB
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Scientific contextConjugation of ubiquitin to proteins (ubiquitination) plays a fundamental role in the regulation of cellular function through biological events involving, amongst others, cell cycle, differentiation, immune responses, DNA repair, chromatin structure, and apoptosis. The ubiquitin signaling system includes a large family of cysteine proteases known as deubiquitinating enzymes (DUBs) that are responsible for the removal of ubiquitin from modified proteins. This regulatory process allows optimal levels of cellular ubiquitin to be maintained by recycling ubiquitin attached to inappropriate targets, removing and disassembling polyubiquitin chains, and processing proteins prior to their degradation by the proteasome. DUBs in general exhibit a wide range of substrate, ubiquitination type (mono- or poly-ubiquitination) and polyubiquitin chain linkage specificities and can be partnered with various interacting proteins to facilitate increased diversity in specificity and DUB activation. DUBs have been implicated in a number of human diseases including various forms of cancer and neurodegeneration. As such they are attractive targets for potential therapeutic intervention via the development of suitable inhibitors and modulators.
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Primary research areaCell Signaling, Post-Translational Modifications, Ubiquitination, Epigenetics, Cancer, Apoptosis, Autophagy, Neuroscience, Neurodegeneration, Cardiovascular System
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CategoryAssay Activity Kits
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Other nameDeubiquitinating enzyme Activity Kit
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Brand nameStressXpress®
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Detection systemFluorometric Assay
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Assay formatContinuous Kinetic Enzyme Activity Assay
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Main intended usageActivity kit used to screen potential inhibitors and activators against DUB enzymes, and to assess and evaluate performance of DUB activity in purified protein samples. It can also be used to optimize assays for specific DUBs to facilitate their use in HTS, and demonsrate that novel DUB enzymes have ubiquitin-AMC processing activity.
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Brief protocolThe DUB Activity kit facilitates the rapid, robust measurement of deubiquitinating enzyme activity in vitro. The kit utilises high purity, fluorogenic substrate ubiquitin-AMC together with suitable calibration standards and controls for the accurate and sensitive assessment of DUB activity. Continuous kinetic or end-point assays can be performed in 96-well plate format for multi-sample analysis. Contains sufficient materials for one full 96-well plate assay set-up to be run.
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Kit contents10x StressXpress® DUB assay buffer, Control DUB enzyme (USP2 catalytic domain), Ubiquitin-AMC, StressXpress® 96 well assay plate, StressXpress® AMC standard
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Assay precisionContact us
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Assay cross reactivitySpecies Independent
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Samples to be used with this kitPurified Proteins
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Maximum samples to be used with this kit40 samples in duplicate
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Assay duration30-90 minutes
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Kit s sensitivitySee product datasheet or feel free to contact us
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Assay detection limitSee product datasheet or feel free to contact us
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Storage recommendations-80
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Shipping recommendationsDry Ice
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Bibliography1. Komander, D. & Rape, M. Annual Review of Biochemistry 81, 203–229 (2012). 2. Ciechanover, A. Biochimica et biophysica acta 1824, 3–13 (2012). 3. Clague, M. J., Coulson, J. M. & Urbé, S. Journal of cell science 125, 277–86 (2012). 4. Komander, D., Clague, M. J. & Urbé, S. Nature reviews. Molecular cell biology 10, 550–63 (2009). 5. Fraile, J. M., Quesada, V., Rodríguez, D., Freije, J. M. P. & López-Otín, C. Oncogene 31, 2373–88 (2012). 6. Kessler, B. M. Current opinion in chemical biology 17, 59-65 (2013). 7. Bilguvar, K. et al. Proceedings of the National Academy of Sciences of the United States of America 110, 3489–94 (2013).
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Release date1-Jun-2014
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PubMed numberRefer to PubMed
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Tested applicationsNo data available / currently testing
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Tested reactivityNo data available / currently testing
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Resources available upon requestKit Booklet, MSDS
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Product image link
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Product image legendContinuous kinetic enzyme activity assay showing the time course of control DUB enzyme (USP2 CD) activity with Ub-AMC substrate using StressXpress® DUB Activity Assay kit – Control DUB (10nM) was incubated with 500nM Ub-AMC in DUB assay buffer for 15 minutes at room temperature alongside a substrate only (blank) control reaction. Fluorescence measurements (RFU) were taken at 30 second intervals and plotted vs. time | AMC Standard Curve for StressXpress® DUB Activity Assay Kit. Representative plot of fluorescence measurements (RFU) of serial dilutions of AMC Standard at recommended concentrations. Enzyme Activity Graph of the Continuous Kinetic Assay for the DUB Activity Kit StressXpress - SKT-136 | Graph of the AMC Typical Standard Curve for the DUB Activity Kit StressXpress - SKT-136
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WarningsNon-hazardous
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Country of productionCanada
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Total weight kg2
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Net weight g300
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Gene target
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Short nameDUB Activity Kit
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Alternative nameDUB Activity reagent
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Alternative techniquekits
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Alternative to gene targetv-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog, C-Kit and CD117 and PBT and SCFR, KIT and IDBG-18980 and ENSG00000157404 and 3815, transferase activity, Extracellular, Kit and IDBG-172083 and ENSMUSG00000005672 and 16590, KIT and IDBG-642326 and ENSBTAG00000002699 and 280832
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