In-gel proteasome assay to determine the activity, amount, and composition of proteasome complexes from mammalian cells or tissues

Summary: This protocol describes an easy and reliable in-gel proteasome assay to quantify the activity and composition of different proteasome complexes in cells and tissues. The assay works well with limited amounts of total cell protein lysates. Although this assay is optimized specifically for th...

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Bibliographic Details
Main Authors: Ayse Seda Yazgili, Thomas Meul, Vanessa Welk, Nora Semren, Ilona E. Kammerl, Silke Meiners
Format: Article
Language:English
Published: Elsevier 2021-06-01
Series:STAR Protocols
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166721002331
Description
Summary:Summary: This protocol describes an easy and reliable in-gel proteasome assay to quantify the activity and composition of different proteasome complexes in cells and tissues. The assay works well with limited amounts of total cell protein lysates. Although this assay is optimized specifically for the proteasome chymotrypsin-like activity, it can be expanded to other proteasome activities as well. Using antibodies that detect distinct proteasome subunits or regulators, we can determine the composition and relative quantity of active proteasome complexes.For complete details on the use and execution of this protocol, please refer to Meul et al. (2020).
ISSN:2666-1667