| 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: |
Assay Kits |
| Storage Temp: |
-20ºC |
| Species Reactivity: |
ALL |
| Data Sheet: |
PLEASE CLICK HERE TO DOWNLOAD |
| Application Details: |
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.
|
| 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: |
Activity Kits |
| 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). |
| Country of Origin: |
Canada |
| Assay Type: |
Fluorometric Assay |
| Assay Type Abbreviation: |
Continuous Kinetic Assay |
| Platform: |
Enzyme Activity |
| Utility: |
Microplate |
| Assay Overview: |
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.
|
| 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 |
|
