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在微载体培养珠上分离细胞质膜

Isolation of cell plasma membranes on microcarrier culture beads.

作者信息

Gotlib L J

出版信息

Biochim Biophys Acta. 1982 Feb 8;685(1):21-6. doi: 10.1016/0005-2736(82)90029-3.

Abstract

A simple, efficient method for the purification of plasma membranes from cultured cells is presented. Membrane purification is effected by attachment of viable cells to commercially available microcarrier culture beads, followed by lysis of the cells, agitation on a vortex mixer and sonication. Optimal conditions for each step of the procedure are described. Enzyme markers from plasma membranes are purified 10-20-fold relative to whole cell homogenates while internal membrane markers are depleted 10-20-fold.

摘要

本文介绍了一种从培养细胞中纯化质膜的简单、高效方法。膜纯化通过将活细胞附着于市售的微载体培养珠上,然后裂解细胞、在涡旋混合器上搅拌并超声处理来实现。描述了该程序每个步骤的最佳条件。相对于全细胞匀浆,质膜的酶标志物纯化了10至20倍,而内膜标志物减少了10至20倍。

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