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Proteasome glo chymotrypsin like assay kit

Manufactured by Promega
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Sourced in United States, Australia

The Proteasome-Glo Chymotrypsin-Like Assay kit is a luminescent-based assay designed to measure the chymotrypsin-like activity of the proteasome in cell lysates or purified proteasome samples. The kit provides a specific substrate and luminescent detection reagent to quantify proteasome activity.

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Lab products found in correlation

Plate reading luminometer, by Agilent Technologies (2 mentions) Gm2000, by Promega (1 mentions) Spark multimode microplate, by Tecan (1 mentions)

Close spelling variants of this product

Proteasome-GloTM Chymotrypsin-like Cell-Based Assays kit Proteasome-Glo assay Proteasome-Glo kit Proteasome-Glo Chymotrypsin

4 protocols using proteasome glo chymotrypsin like assay kit

1

Proteasome Activity Assay in Liver

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The proteasome chymotrypsin-like activity of liver was detected using Proteasome-Glo Chymotrypsin-Like Assay kit (Promega) following the manufacturer's instruction. In brief, protein concentration of liver lysates from control mice and fasted mice was measured by Bradford assay. Equal amount of liver extract was diluted with polysome buffer to a final volume of 100 μL and loaded to a white 96-well plate. 100 μL of Proteasome-Glu reagent was added to each sample and incubated at room temperature for 10 mins. Luminescence was measured by a plate-reading luminometer (BioTek).
Gao X., Wan J., Liu B., Ma M., Shen B, & Qian S.B. (2014). Quantitative profiling of initiating ribosomes in vivo. Nature methods, 12(2), 147-153.
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2

Proteasome Activity Measurement Assay

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The Proteasome-Glo™ Chymotrypsin-Like Assay Kit (G8622; Promega, Madison, WI, USA) was used to measure proteasome activity. Cells were collected and homogenized; equal concentrations of proteins were added to Proteasome-Glo™ buffer, Suc-LLVY-Glo™ substrate, and luciferin detection reagent to separate groups; and luminescence was detected with a luminometer (GM2000; Promega).
Chao C.C., Huang C.L., Cheng J.J., Chiou C.T., Lee I.J., Yang Y.C., Hsu T.H., Yei C.E., Lin P.Y., Chen J.J, & Huang N.K. (2022). SRT1720 as an SIRT1 activator for alleviating paraquat-induced models of Parkinson's disease. Redox Biology, 58, 102534.
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3

20S Proteasome Inhibition Assay

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The 20S proteasome inhibition assay was performed using the Proteasome-Glo™ Chymotrypsin-Like Assay kit (Promega, Alexandria, NSW, Australia). Suc-LLVY-aminoluciferin is a substate that can be cleaved by the 20S proteasome, release the luciferase (aminoluciferin) and allow the luciferase reaction to produce light. Gradient concentration of tested compounds and the purified 20S proteasome enzyme (R&D systems, Minneapolis, MN, USA) were successively added into a transparent 96-well plate and mixed thoroughly. Then, 50 µL of a Suc-LLVY-AMC (10 μM) reagent was added. Gently mix contents with a plate shaker at 300 to 500 rpm for 30 s. Incubate at room temperature for an hour. The luminescence was recorded via the Tecan Spark multimode microplate.
Zhang J., Sun S., Liu J., Zhang L., Guo D., Zhang N., Zhao J., Kong D., Xu T., Wang X., Xu W., Li X, & Jiang Y. (2023). Discovery of a Novel Ubenimex Derivative as a First-in-Class Dual CD13/Proteasome Inhibitor for the Treatment of Cancer. Molecules, 28(17), 6343.
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4

Proteasome Activity Assay in Liver

Check if the same lab product or an alternative is used in the 5 most similar protocols
The proteasome chymotrypsin-like activity of liver was detected using Proteasome-Glo Chymotrypsin-Like Assay kit (Promega) following the manufacturer's instruction. In brief, protein concentration of liver lysates from control mice and fasted mice was measured by Bradford assay. Equal amount of liver extract was diluted with polysome buffer to a final volume of 100 μL and loaded to a white 96-well plate. 100 μL of Proteasome-Glu reagent was added to each sample and incubated at room temperature for 10 mins. Luminescence was measured by a plate-reading luminometer (BioTek).
Gao X., Wan J., Liu B., Ma M., Shen B, & Qian S.B. (2014). Quantitative profiling of initiating ribosomes in vivo. Nature methods, 12(2), 147-153.
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