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20S Proteasome Activity Kit StressMarq SKT-133-96 96 well £366.00
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Product Detail: Fluorometric detection of purified 20S proteasome activity
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: Activity Kits
Storage Temp: 4ºC
Species Reactivity: ALL
Data Sheet: PLEASE CLICK HERE TO DOWNLOAD
Target: 20S Proteasome
Research Area(s): Cancer | Apoptosis | Cell Signaling | Post-translational Modifications | Neuroscience | Neurodegeneration | Cardiovascular System
Alternative Name(s): Chymotrypsin-like (?5) Activity Kit
Category: Assay Kits
Species Reactivity Full Name: Species Independent
Shipping Temperature: Blue Ice
References: 1. Saeki, Y. & Tanaka, K. Methods in molecular biology (Clifton, N.J.) 832, 315–37 (2012).
2. Frankland-Searby, S. & Bhaumik, S. R. Biochimica et biophysica acta 1825, 64–76 (2012).
3. Basler, M., Kirk, C. J. & Groettrup, M. Current opinion in immunology 1–7 (2012).
4. Löw, P. General and comparative endocrinology 172, 39–43 (2011).
5. Dennissen, F. J. a, Kholod, N. & Van Leeuwen, F. W. Progress in neurobiology 96, 190–207 (2012).
6. Geng, F., Wenzel, S. & Tansey, W. P. Annual review of biochemistry 81, 177–201 (2012).
Launch Date: 1/Jun/2014
Tarriff Code: 3822.00.1090
ADR Code: Non-hazardous
UN Code for transport: Non-hazardous
Country of Origin: Canada
Assay Type: Continuous Kinetic Assay
Assay Type Abbreviation: Enzyme Activity
Platform: Microplate
Utility: 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.
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.
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
Cite this Product: 20S Proteasome Activity Kit (StressMarq Biosciences, Canada, Cat # SKT-133-96)
Trademark: StressXpress®