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| | Proteasome Activity Assay |
| Description: | Activity assays were carried out in a 200 uL reaction volume. Different concentrations of test compounds were added to a black flat/clear bottom 96-well plate containing 1 nM of either human constitutive 20S proteasome or 26S proteasome, in 50 mM Tris-HCl at pH 7.5 and allowed to sit for 10 min at RT. Fluorogenic substrates were then added and the enzymatic activity measured at 37 °C on a SpectraMax M5e spectrometer by measuring the change in fluorescence unit per minute for 1 h at 380−460 nm. The fluorescence units for the vehicle control were set at a 100%, and the ratio of drug-treated sample set to that of vehicle control was used to calculate the fold change in enzymatic activity. |
| Affinity data for this assay | |
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