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

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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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:
Antibody
Cell Biology
Molecular Biology
Protein Biochemistry
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166721002331
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spelling doaj-9909539c0bd24599b3ba50ef6e1e7f802021-06-21T04:25:32ZengElsevierSTAR Protocols2666-16672021-06-0122100526In-gel proteasome assay to determine the activity, amount, and composition of proteasome complexes from mammalian cells or tissuesAyse Seda Yazgili0Thomas Meul1Vanessa Welk2Nora Semren3Ilona E. Kammerl4Silke Meiners5Comprehensive Pneumology Center (CPC), University Hospital of the Ludwig-Maximilians-University (LMU) and Helmholtz Zentrum München, Member of the German Center for Lung Research (DZL), Max-Lebsche Platz 31, 81377 Munich, Germany; Corresponding authorComprehensive Pneumology Center (CPC), University Hospital of the Ludwig-Maximilians-University (LMU) and Helmholtz Zentrum München, Member of the German Center for Lung Research (DZL), Max-Lebsche Platz 31, 81377 Munich, GermanyComprehensive Pneumology Center (CPC), University Hospital of the Ludwig-Maximilians-University (LMU) and Helmholtz Zentrum München, Member of the German Center for Lung Research (DZL), Max-Lebsche Platz 31, 81377 Munich, GermanyComprehensive Pneumology Center (CPC), University Hospital of the Ludwig-Maximilians-University (LMU) and Helmholtz Zentrum München, Member of the German Center for Lung Research (DZL), Max-Lebsche Platz 31, 81377 Munich, GermanyComprehensive Pneumology Center (CPC), University Hospital of the Ludwig-Maximilians-University (LMU) and Helmholtz Zentrum München, Member of the German Center for Lung Research (DZL), Max-Lebsche Platz 31, 81377 Munich, GermanyComprehensive Pneumology Center (CPC), University Hospital of the Ludwig-Maximilians-University (LMU) and Helmholtz Zentrum München, Member of the German Center for Lung Research (DZL), Max-Lebsche Platz 31, 81377 Munich, Germany; Corresponding authorSummary: 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).http://www.sciencedirect.com/science/article/pii/S2666166721002331AntibodyCell BiologyMolecular BiologyProtein Biochemistry
collection DOAJ
language English
format Article
sources DOAJ
author Ayse Seda Yazgili
Thomas Meul
Vanessa Welk
Nora Semren
Ilona E. Kammerl
Silke Meiners
spellingShingle Ayse Seda Yazgili
Thomas Meul
Vanessa Welk
Nora Semren
Ilona E. Kammerl
Silke Meiners
In-gel proteasome assay to determine the activity, amount, and composition of proteasome complexes from mammalian cells or tissues
STAR Protocols
Antibody
Cell Biology
Molecular Biology
Protein Biochemistry
author_facet Ayse Seda Yazgili
Thomas Meul
Vanessa Welk
Nora Semren
Ilona E. Kammerl
Silke Meiners
author_sort Ayse Seda Yazgili
title In-gel proteasome assay to determine the activity, amount, and composition of proteasome complexes from mammalian cells or tissues
title_short In-gel proteasome assay to determine the activity, amount, and composition of proteasome complexes from mammalian cells or tissues
title_full In-gel proteasome assay to determine the activity, amount, and composition of proteasome complexes from mammalian cells or tissues
title_fullStr In-gel proteasome assay to determine the activity, amount, and composition of proteasome complexes from mammalian cells or tissues
title_full_unstemmed In-gel proteasome assay to determine the activity, amount, and composition of proteasome complexes from mammalian cells or tissues
title_sort in-gel proteasome assay to determine the activity, amount, and composition of proteasome complexes from mammalian cells or tissues
publisher Elsevier
series STAR Protocols
issn 2666-1667
publishDate 2021-06-01
description 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).
topic Antibody
Cell Biology
Molecular Biology
Protein Biochemistry
url http://www.sciencedirect.com/science/article/pii/S2666166721002331
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