Journal
NATURE PROTOCOLS
Volume 4, Issue 11, Pages 1582-1590Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2009.151
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Funding
- Polish State Committee for Scientific Research [N301 092 32/3407]
- Polish Mitochondrial Network
- AIRC
- UMDF
- PRRIITT
- FISM
- Telethon [GGP09128]
- University of Ferrara
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Many cellular processes require the proper cooperation between mitochondria and the endoplasmic reticulum (ERER). Several recent works show that their functional interactions rely on dynamic structural contacts between both organelles. Such contacts, called mitochondria-associated membranes (MAMs), are crucial for the synthesis and intracellular transport of phospholipids, as well as for intracellular Ca2+ signaling and for the determination of mitochondrial structure. Although several techniques are available to isolate mitochondria, only few are specifically tuned to the isolation of MAM, containing unique regions of ERER membranes attached to the outer mitochondrial membrane and mitochondria without contamination from other organelles (i.e., pure mitochondria). Here we provide optimized protocols to isolate these fractions from tissues and cells. These procedures require 4-5 h and can be easily modified and adapted to different tissues and cell types.
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