Proteasome 20S Activity Assay Kit (Fluorometric)
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(4 Publications)
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Functional Studies - Proteasome 20S Activity Assay Kit (Fluorometric) (AB112154)
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 = 490/525.
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Product details
Proteasome 20S Activity Assay Kit ab112154 is a homogeneous fluorescent assay that measures the chymotrypsin-like protease activity associated with the proteasome complex in cultured cells. ab112154 uses LLVY-R110 as a fluorogenic indicator for proteasome activities. Cleavage of LLVY-R110 by the proteasome generates strongly green fluorescent R110 that is monitored fluorometrically at 520-530 nm with excitation at 480-500 nm. The kit provides all the essential components with an optimized assay protocol. The assay is robust, and can be readily adapted for high-throughput assays to evaluate the proteasome activities or screen inhibitors in cultured cells or in solution. The assay can be performed in a convenient 96-well and 384-well fluorescence microtiter-plate format. Visit our for tips and troubleshooting.
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Biological function summary
The proteasome 20S operates within a larger proteolytic complex known as the 26S proteasome essential for ATP-dependent protein degradation. The 20S core decomposes substrates into peptides through its proteolytic activity facilitated by its catalytic subunits. The proteasome activity is important for cellular processes such as cell cycle regulation apoptosis and DNA repair. This activity can be measured using a proteasome activity assay where fluorogenic assays are often used to quantify its enzymatic function.
Pathways
Proteasome 20S contributes significantly to proteostasis and the regulation of protein turnover. It involves itself in the ubiquitin-proteasome pathway which plays a pivotal role in cellular response to oxidative stress and is important for the NF-kB pathway. Its activity interacts with various proteins such as ubiquitin ligases and deubiquitinating enzymes which help target proteins for degradation. These interactions ensure timely removal of proteins preventing toxic accumulation and ensuring proper cellular functioning.
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Publications (4)
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Methods in molecular biology (Clifton, N.J.) 2712:135-148 PubMed37578702
2023
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Human reproduction (Oxford, England) 37:2095-2112 PubMed35856882
2022
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International journal of molecular medicine 47: PubMed33537802
2021
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FEBS open bio 6:1085-1092 PubMed27833849
2016
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