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Fluorogenic substrate proteasome activity assay
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HCT-8 cells were collected and resuspended in 200 μL of proteasome activity assay buffer (115 mM NaCl, 1 mM KH2PO4, 5 mM CaCl2, 1.2 mM MgSO4, 25 mM sodium HEPES buffer (pH 7.4)) containing 62.5 mM Suc-Leu-Leu-Val-Tyr-AMC (Suc-LLVY-AMC; Enzo Life Sciences, Farmingdale, NY, USA). The same quantity of cell lysate was incubated at 37 °C for 1 h and then measured at 360 nm/460 nm using a Victor3 Multilabel Plate Reader (Perkin Elmer, Waltham, MA, USA).
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