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

凝胶内蛋白酶体分析法测定哺乳动物细胞或组织中蛋白酶体复合物的活性、数量和组成

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作者:Ayse Seda Yazgili ,Thomas Meul ,Vanessa Welk ,Nora Semren ,Ilona E Kammerl ,Silke Meiners

Abstract

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). Keywords: Antibody; Cell Biology; Molecular Biology; Protein Biochemistry.

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