DUB Activity Kit

  • Catalog number
    SKT-136-96
  • Price
    Please ask
  • Size
    96 well
  • Stock availability
    In Stock
  • Kit s description
    Fluorometric detection of DUB activity
  • Protein target
    DUB
  • Scientific context
    Conjugation 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.
  • Primary research area
    Cell Signaling, Post-Translational Modifications, Ubiquitination, Epigenetics, Cancer, Apoptosis, Autophagy, Neuroscience, Neurodegeneration, Cardiovascular System
  • Category
    Assay Activity Kits
  • Other name
    Deubiquitinating enzyme Activity Kit
  • Brand name
    StressXpress®
  • Detection system
    Fluorometric Assay
  • Assay format
    Continuous Kinetic Enzyme Activity Assay
  • Main intended usage
    Activity 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.
  • Brief protocol
    The 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.
  • Kit contents
    10x StressXpress® DUB assay buffer, Control DUB enzyme (USP2 catalytic domain), Ubiquitin-AMC, StressXpress® 96 well assay plate, StressXpress® AMC standard
  • Assay precision
    Contact us
  • Assay cross reactivity
    Species Independent
  • Samples to be used with this kit
    Purified Proteins
  • Maximum samples to be used with this kit
    40 samples in duplicate
  • Assay duration
    30-90 minutes
  • Kit s sensitivity
    See product datasheet or feel free to contact us
  • Assay detection limit
    See product datasheet or feel free to contact us
  • Storage recommendations
    -80
  • Shipping recommendations
    Dry Ice
  • Bibliography
    1. 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).
  • Release date
    1-Jun-2014
  • PubMed number
    Refer to PubMed
  • Tested applications
    No data available / currently testing
  • Tested reactivity
    No data available / currently testing
  • Resources available upon request
    Kit Booklet, MSDS
  • Product image link
  • Product image legend
    Continuous 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
  • Warnings
    Non-hazardous
  • Country of production
    Canada
  • Total weight kg
    2
  • Net weight g
    300
  • Gene target
    DUB   Activity   Kit  
  • Short name
    DUB Activity Kit
  • Alternative name
    DUB Activity reagent
  • Alternative technique
    kits
  • Alternative to gene target
    v-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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