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
Descripción
Sumario: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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