| Background Info: |
Proteasomes are non-lysosomal proteolytic complexes localised primarily in the cytoplasm and in the nucleus of eukaryotic cells. They are responsible for the ubiquitin-mediated degradation of short half-life proteins and peptides that are involved in essential cellular processes including cell-cycle regulation, apoptosis and transcriptional regulation, innate immunity and antigen processing, and in the removal of redundant or damaged proteins. As such protein degradation by the ubiquitin-proteasome pathway has a major regulatory function for proliferation activity and survival of both normal and malignant cells, and its dysfunction has been implicated in a wide range of other disease processes including neurodegenerative, cardiovascular and metabolic disorders.
The 26S proteasome structure is composed of a 20S proteasome catalytic core complex and one or two 19S (PA700) regulatory subcomplexes. The 20S core comprises two copies of 14 subunits (7 alpha subunits and 7 beta subunits) arranged in a ?7?7?7?7 cylindrical array. Proteolytic activities are determined by the ?1 (caspase-like), ?2 (trypsin-like) and ?5 (chymotrypsin-like) subunits, access to which is guarded by the ?-subunits. The 19S regulatory unit consists of six ATPase and at least ten non-ATPase subunits that are required for ubiquitinated protein binding, deubiquitination, substrate unfolding and translocation to the 20S catalytic core.
Varying catalytic subunit composition (?1, ?1i; ?2, ?2i; ?5, ?5i) results in a variety of possible subtypes from full constitutive proteasome (?1, ?2, ?5) through mixed populations to full inducible / immunoproteasome (?1i, ?2i, ?5i). Alternative regulatory complexes such as the PA200 and 11S proteasome activators confer different substrate specificities and activity compared to the 19S regulator. |
| Product Type: |
Assay Kits |
| Storage Temp: |
4ºC |
| Species Reactivity: |
ALL |
| Applications: |
Activity kit used to screen potential proteasome inhibitors and activators, and to measure and quantify 20S proteasome activity in continuous kinetic and end-point assays.
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| Research Area(s): |
Cancer | Apoptosis | Cell Signaling | Post-translational Modifications | Neuroscience | Neurodegeneration | Cardiovascular System |
| Target: |
20S Proteasome |
| Alternative Name(s): |
Chymotrypsin-like (?5) Activity Kit |
| Category: |
Activity Kits |
| Trademark: |
StressXpress® |
| Species Reactivity Full Name: |
Species Independent |
| References: |
1. Saeki, Y. & Tanaka, K. Methods in molecular biology (Clifton, N.J.) 832, 31537 (2012).
2. Frankland-Searby, S. & Bhaumik, S. R. Biochimica et biophysica acta 1825, 6476 (2012).
3. Basler, M., Kirk, C. J. & Groettrup, M. Current opinion in immunology 17 (2012).
4. Löw, P. General and comparative endocrinology 172, 3943 (2011).
5. Dennissen, F. J. a, Kholod, N. & Van Leeuwen, F. W. Progress in neurobiology 96, 190207 (2012).
6. Geng, F., Wenzel, S. & Tansey, W. P. Annual review of biochemistry 81, 177201 (2012). |
| Tarriff Code: |
3822.00.1090 |
| ADR Code: |
Non-hazardous |
| UN Code for transport: |
Non-hazardous |
| Country of Origin: |
Canada |
| Cite this Product: |
20S Proteasome Activity Kit (StressMarq Biosciences, Canada, Cat # SKT-133-96) |
| Assay Type: |
Continuous Kinetic Assay |
| Assay Type Abbreviation: |
Enzyme Activity |
| Platform: |
Microplate |
| Assay Overview: |
The 20S Proteasome Activity kit facilitates the rapid, robust measurement of proteasome activity in vitro. The kit utilises high purity, fluorogenic substrate Suc-LLVY-AMC together with suitable calibration standards and controls for the accurate and sensitive assessment of proteasome chymotrypsin-like (?5) activity. Continuous kinetic or end-point assays can be performed in micro-cuvettes or in 96-well plate format for multi-sample analysis. Contains sufficient materials for one full 96-well plate assay or up to 100x 50µL micro-cuvette based assays to be run.
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| Kit Components: |
10x StressXpress® proteasome assay buffer | 20S proteasome | MG132 (control inhibitor) | StressXpress® 96 well assay plate | StressXpress® AMC standard | Suc-LLVY-AMC (chymotrypsin-like substrate) |
| Sample Type(s): |
Purified Proteins |
| Number of Samples: |
40 samples in duplicate using the plate format, or 100x 50µL micro-cuvette based assays |
| Incubation Time: |
15-30 minutes |
| Other relevant information: |
Fluorometric Assay |
| Field of Use: |
Not for use in humans. Not for use in diagnostics or therapeutics. For in vitro research use only. |
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