title: Isolation of mitochondria-associated ER membranes (MAMs) and glycosphingolipid-enriched microdomains (GEMs) from brain tissues and neuronal cells.
abstract: Subcellular fractionation is a valuable procedure in cell biology to separate and purify various subcellular constituents from one another, i.e., nucleus, cytosol, membranes/organelles, and cytoskeleton. The procedure relies on the use of differential centrifugation of cell and tissue homogenates, but additional purification steps now permit the isolation of inter-organellar membrane contact sites. Here, we outline a protocol tailored for the isolation of mitochondria, mitochondria-associated ER membranes (MAMs) and glycosphingolipid-enriched microdomains (GEMs) from the adult mouse brain, primary neurospheres, and murine embryonic fibroblasts (MEFs).
authors: Department of Genetics, St. Jude Children Research Hospital, 262 Danny Thomas Pl., Memphis, TN, 38105, USA.
authors: Ida
authors: I
authors: Annunziata
authors: Annette
authors: A
authors: Patterson
authors: Alessandra
authors: A
authors: d'Azzo
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism*; D002458:Cell Fractionation / Q000379:methods*; D004721:Endoplasmic Reticulum / Q000737:chemistry / Q000378:metabolism*; D006028:Glycosphingolipids / Q000737:chemistry / Q000378:metabolism*; D007425:Intracellular Membranes / Q000737:chemistry / Q000378:metabolism*; D021962:Membrane Microdomains / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008928:Mitochondria / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D006028:Glycosphingolipids