Isolation of mitochondria-associated ER membranes (MAMs) and glycosphingolipid-enriched microdomains (GEMs) from brain tissues and neuronal cells.
Isolation of mitochondria-associated ER membranes (MAMs) and glycosphingolipid-enriched microdomains (GEMs) from brain tissues and neuronal cells.
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DOI：
10.1007/978-1-4939-2257-4_3
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发表时间：
2015-01-01
期刊：
影响因子：
--
通讯作者：
d'Azzo, Alessandra
中科院分区：
文献类型：
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作者：
Annunziata, Ida;Patterson, Annette;d'Azzo, Alessandra
亚细胞分级分离在细胞生物学中是一种有价值的方法，用于将各种亚细胞成分彼此分离和纯化，即，细胞核、细胞溶质、膜/细胞器和细胞骨架。该程序依赖于使用差速离心的细胞和组织匀浆，但额外的纯化步骤，现在允许的细胞器间膜接触部位的隔离。在这里，我们概述了一个协议定制的线粒体，线粒体相关的ER膜（MAMs）和鞘糖脂富集的微结构域（GEM）从成年小鼠大脑，初级神经球，和小鼠胚胎成纤维细胞（MEFs）的分离。
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).