Key features and details
- Assay type: Quantitative
- Detection method: Fluorescent
- Platform: Microplate reader
- Sample type: Adherent cells, Suspension cells
Product nameProteasome 20S Activity Assay Kit (Fluorometric)
Sample typeAdherent cells, Suspension cells
Species reactivityReacts with: Mammals, Other species
ab112154 Proteasome 20S Activity Assay Kit 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.
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Storage instructionsStore at -20°C. Please refer to protocols.
Components Identifier 1 x 96 tests Assay Buffer 1 x 10ml DMSO 1 x 100µl Proteasome LLVY-R110 Substrate Component A 1 vial
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.
ab112154 has been referenced in 2 publications.
- Bi L et al. Paris saponin H inhibits the proliferation of glioma cells through the A1 and A3 adenosine receptor-mediated pathway. Int J Mol Med 47:N/A (2021). PubMed: 33537802
- Wu T et al. Rrp15 affects cell cycle, proliferation, and apoptosis in NIH3T3 cells. FEBS Open Bio 6:1085-1092 (2016). PubMed: 27833849