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Cold Spring Harb Protoc. 2014 Oct 1;2014(10):pdb.prot080002. doi: 10.1101/pdb.prot080002.

Isolation of mitochondria from tissue culture cells.

Author information

1
Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia 20147-2408;
2
Department of Pathology, Yale University School of Medicine, New Haven, Connecticut 06520-8023; Department of Genetics, Yale University School of Medicine, New Haven, Connecticut 06520-8023.

Abstract

The number of mitochondria per cell varies substantially from cell line to cell line. For example, human HeLa cells contain at least twice as many mitochondria as smaller mouse L cells. This protocol starts with a washed cell pellet of 1-2 mL derived from ∼10⁹ cells grown in culture. The cells are swollen in a hypotonic buffer and ruptured with a Dounce or Potter-Elvehjem homogenizer using a tight-fitting pestle, and mitochondria are isolated by differential centrifugation.

PMID:
25275104
DOI:
10.1101/pdb.prot080002
[Indexed for MEDLINE]

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