从分离的神经元制备质膜。
Preparation of plasma membrane from isolated neurons.
作者信息
Henn F A, Hansson H A, Hamberger A
出版信息
J Cell Biol. 1972 Jun;53(3):654-61. doi: 10.1083/jcb.53.3.654.
Abstract
A bulk fraction enriched with respect to neuronal cell bodies was used as starting material for the isolation of neuronal plasma membrane The cells were gently homogenized in isotonic sucrose and a crude membrane containing fraction sedimented at 3000 g. Subsequently, the membrane fraction was purified on a discontinuous sucrose density gradient between 35% and 25 5% sucrose (w/w). Enzymatic analyses showed a 4-5-fold enrichment in plasma membrane markers, and a 10-15% contamination of mitochondrial and microsomal material. Electron micrographs of the membrane fraction confirmed the enzymatic data Fragmented membranes were found, mainly in vesicular form No ribosomes, but a few mitochondria and some multilamellar membranes were seen
摘要
富含神经元细胞体的大量部分被用作分离神经元质膜的起始材料。细胞在等渗蔗糖中轻轻匀浆，含有粗膜的部分在3000g下沉淀。随后，膜部分在35%至25.5%蔗糖（w/w）的不连续蔗糖密度梯度上纯化。酶分析表明质膜标志物富集了4至5倍，线粒体和微粒体物质的污染为10%至15%。膜部分的电子显微照片证实了酶学数据。发现了破碎的膜，主要呈囊泡形式。未见核糖体，但可见一些线粒体和一些多层膜。
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