Product Name | DUB Activity Kit |
Description |
Fluorometric detection of DUB activity |
Species Reactivity | Species Independent |
Platform | Microplate |
Sample Types | Purified Proteins |
Detection Method | Fluorometric Assay |
Assay Type | Continuous Kinetic Assay |
Utility | 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. |
Incubation Time | 30-90 minutes |
Number of Samples | 40 samples in duplicate |
Other Resources | Kit Booklet , MSDS |
Field of Use | Not for use in humans. Not for use in diagnostics or therapeutics. For in vitro research use only. |
Storage Temperature | -80 | ||||||||||||||||||
Shipping Temperature | Dry Ice | ||||||||||||||||||
Product Type | Activity Kits | ||||||||||||||||||
Assay Overview | 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 Overview |
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Cite This Product | DUB Activity Kit (StressMarq Biosciences Inc., Victoria BC CANADA, Catalog # SKT-136) |
Alternative Names | Deubiquitinating enzyme Activity Kit |
Research Areas | Apoptosis, Autophagy, Cancer, Cardiovascular System, Cell Signaling, Neurodegeneration, Neuroscience, Post-translational Modifications, Ubiquitination |
Scientific Background | 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. |
References |
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). |
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
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