After the above treatment, the 96-well plates were equilibrated at room temperature for 10 min before adding the reagents to detect immunoproteasome activities. To detect chymotrypsin-like, trypsin-like and caspase-like enzymatic activities, an equal volume of the Proteasome-Glo Cell-Based Reagent (Promega, Madison, WI, USA) was added in each well according to the manufacturer’s instruction. The plates were incubated at room temperature for 10 min in the dark and the relative luminescence units (RLU) of assays were determined in the LUMIstar Omega microplate luminometer (BMG Labtech, Ortenberg, Germany) according to the manufacturer’s directions.
Proteasome glo cell based reagent
Manufactured by Promega
Sourced in United States
The Proteasome-Glo Cell-Based Reagent is a luminescent assay for detecting and quantifying proteasome activity in live cells. It provides a simple, sensitive, and reproducible method for measuring proteasome function.
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Lab products found in correlation
2 protocols using proteasome glo cell based reagent
1
Proteasome Activity Profiling in Cells
2
Proteasome Activity Assay in HEK293 Cells
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HEK293 cells were plated equally (10,000 cells per well) in 96-well white-walled plates. The wells were subjected to amino acid starvation conditions for various time points (0, 0.5, 1, 2, and 3 h, respectively). Proteasome-Glo Cell-Based Reagent (Promega, catalog # G8660) was prepared as per manufacturer's protocol and an equal volume was added to each well. The content of the plate was mixed at 700 RPM for 2 min and then incubated at room temperature for 10 min. Luminescence was read using a Synergy 2 Luminescence Plate Reader (Biotek).
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