Protocol for mitochondrial isolation and sub-cellular localization assay for mitochondrial proteins

Here, we provide a protocol to isolate mitochondria from cultured cells and extract differently located mitochondrial proteins. We detail steps to separate both integral and peripheral membrane proteins from soluble proteins using sonication. We describe the separation of integral membrane proteins...

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Detalles Bibliográficos
Autores principales: Zhou, Danyi, Zhong, Sheng, Han, Xinyu, Liu, Dandan, Fang, Hezhi, Wang, Ya
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9932554/
https://www.ncbi.nlm.nih.gov/pubmed/36853693
http://dx.doi.org/10.1016/j.xpro.2023.102088
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author Zhou, Danyi
Zhong, Sheng
Han, Xinyu
Liu, Dandan
Fang, Hezhi
Wang, Ya
author_facet Zhou, Danyi
Zhong, Sheng
Han, Xinyu
Liu, Dandan
Fang, Hezhi
Wang, Ya
author_sort Zhou, Danyi
collection PubMed
description Here, we provide a protocol to isolate mitochondria from cultured cells and extract differently located mitochondrial proteins. We detail steps to separate both integral and peripheral membrane proteins from soluble proteins using sonication. We describe the separation of integral membrane proteins from the peripheral membrane and soluble proteins using sodium carbonate extraction. Furthermore, we detail the use of proteinase K and Triton X-100 to distinguish outer membrane proteins from mitochondrial proteins.
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spelling pubmed-99325542023-02-17 Protocol for mitochondrial isolation and sub-cellular localization assay for mitochondrial proteins Zhou, Danyi Zhong, Sheng Han, Xinyu Liu, Dandan Fang, Hezhi Wang, Ya STAR Protoc Protocol Here, we provide a protocol to isolate mitochondria from cultured cells and extract differently located mitochondrial proteins. We detail steps to separate both integral and peripheral membrane proteins from soluble proteins using sonication. We describe the separation of integral membrane proteins from the peripheral membrane and soluble proteins using sodium carbonate extraction. Furthermore, we detail the use of proteinase K and Triton X-100 to distinguish outer membrane proteins from mitochondrial proteins. Elsevier 2023-02-03 /pmc/articles/PMC9932554/ /pubmed/36853693 http://dx.doi.org/10.1016/j.xpro.2023.102088 Text en © 2023 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Zhou, Danyi
Zhong, Sheng
Han, Xinyu
Liu, Dandan
Fang, Hezhi
Wang, Ya
Protocol for mitochondrial isolation and sub-cellular localization assay for mitochondrial proteins
title Protocol for mitochondrial isolation and sub-cellular localization assay for mitochondrial proteins
title_full Protocol for mitochondrial isolation and sub-cellular localization assay for mitochondrial proteins
title_fullStr Protocol for mitochondrial isolation and sub-cellular localization assay for mitochondrial proteins
title_full_unstemmed Protocol for mitochondrial isolation and sub-cellular localization assay for mitochondrial proteins
title_short Protocol for mitochondrial isolation and sub-cellular localization assay for mitochondrial proteins
title_sort protocol for mitochondrial isolation and sub-cellular localization assay for mitochondrial proteins
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9932554/
https://www.ncbi.nlm.nih.gov/pubmed/36853693
http://dx.doi.org/10.1016/j.xpro.2023.102088
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