Proteasome 20S Activity Assay Kit
SIGMA/MAK172 - sufficient for 100 fluorometric tests
Synonym: 20S Proteasome Detection Kit
EC Number: 200-664-3
Product Type: Chemical
Proteasome 20S Activity Assay Kit, Cat. No. MAK172: Detection of Proteasome Activity in Jurkat cells. Jurkat cells were seeded on the same day at 500,000 cells/90 μL/well in a 96-well black wall/clear bottom plate. The cells were treated with or without 50 mM H2O2 for 30 minutes. The proteasome assay loading solution (100 μL/well) was added and incubated in a 5% CO2, 37 °C incubator for 3 hours. The fluorescence intensity was measured at Ex/Em = 490/525nm.
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| detection method | fluorometric |
| relevant disease(s) | endocrinological disorders, diabetes; neurological disorders; cancer |
| storage temp. | −20°C |
| usage | sufficient for 100 fluorometric tests |
| Application: | Suitable for measuring the chymotrypsin-like protease activity associated with the proteasome complex either in cultured cells or cell lysates. |
| Biochem/physiol Actions: | The fluorometric proteasome 20S assay kit is a homogeneous fluorescent assay that measures the chymotrypsin-like protease activity associated with the proteasome complex in cultured cells. This kit uses LLVY-R110 as a fluorogenic indicator for proteasome activities. Cleavage of LLVY-R110 by proteasome generates strongly green fluorescent R110 that is monitored fluorimetrically at 520-530 nm with excitation at 480-500 nm. The assay is robust, and can be readily adapted for high-throughput assays to evaluate the proteasome activities or screen the inhibitors in cultured cells or in solution. The assay can be performed in a convenient 96-well and 384-well fluorescence microtiter-plate format. |
| Features and Benefits: | Compatible with high-throughput handling systems. |
| General description: | Protein degradation occurs most commonly through two major mechanisms, lysosomal degradation and proteasome mediated degradation. Lysosomal degradation occurs through proteolytic enzyme activity and is fairly non-specific. Proteasomal degradation is targeted for a specific protein through the ubiquitination of the target protein. The target protein is typically poly-ubiquitinated and activates the 26S proteasome toward proteolysis. The proteasome 26S is the most common form of the proteasome and is an ATP-dependent proteolytic complex, which contains one 20S (700-kDa) core particle structure and two 19S (700-kDa) regulatory caps. The proteasome is responsible for the recycling of proteins and maintenance of the balance of protein synthesis and degradation. The proteasome is involved with apoptosis, DNA repair, endocytosis, and cell cycle and has been implicated in disease states such as certain types of cancer, diabetes mellitus, and Alzheimer′s disease among others making it the target for drug discovery investigations. |
| RIDADR | NONH for all modes of transport |
| Storage Temp. | −20°C |
| UNSPSC | 12161503 |