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免疫沉淀:使用玻璃珠裂解酵母细胞。

Immunoprecipitation: lysing yeast cells using glass beads.

作者信息

Harlow Ed, Lane David

出版信息

CSH Protoc. 2006 Sep 1;2006(4):pdb.prot4533. doi: 10.1101/pdb.prot4533.

Abstract

INTRODUCTIONFor yeast, the preferred method of lysis is mechanical shearing by vortexing the cells in the presence of glass beads as described in this protocol. This is a powerful lysis method but imparts extensive energy into the lysis procedure and results in considerable denaturation of the samples. For this reason, yeast immunoprecipitations are not as powerful a technique as similar studies in mammalian cells and should not be relied upon for determination of protein properties that require authentic protein structure. The lysate prepared in this procedure is ready for preclearing.

摘要

引言

对于酵母而言,本方案中所述的在玻璃珠存在下通过涡旋细胞进行机械剪切是首选的裂解方法。这是一种强大的裂解方法,但会在裂解过程中输入大量能量,导致样品大量变性。因此,酵母免疫沉淀法不如在哺乳动物细胞中进行的类似研究那样有效,不应依赖它来确定需要真实蛋白质结构的蛋白质特性。本步骤中制备的裂解物可用于预清除。

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