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肾近端小管刷状缘膜的分离

Isolation of renal proximal tubular brush-border membranes.

作者信息

Biber Jürg, Stieger Bruno, Stange Gerti, Murer Heini

机构信息

Institute of Physiology, University Zürich, Winterthurerstrasse 190, 8057 Zürich, Switzerland.

出版信息

Nat Protoc. 2007;2(6):1356-9. doi: 10.1038/nprot.2007.156.

Abstract

This protocol describes a method for the isolation and purification of renal proximal tubular brush-border membranes in high yield and high purity. Based on a different reactivity of the brush-border membrane compared to other cellular membranes with divalent cations, such as Mg2+, purified membrane vesicles can be obtained after a few differential centrifugation steps (within approximately 3 h) that are suitable for in vitro studies, such as transport experiments or protein and lipid analysis.

摘要

本方案描述了一种以高产率和高纯度分离和纯化肾近端小管刷状缘膜的方法。基于刷状缘膜与其他细胞膜相比对二价阳离子(如Mg2+)具有不同的反应性,经过几步差速离心(约3小时内)后可获得纯化的膜囊泡,这些膜囊泡适用于体外研究,如转运实验或蛋白质和脂质分析。

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