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胚胎细胞胞核与胞质分离实验方案,用于研究蛋白穿梭。

Protocol for separation of the nuclear and the cytoplasmic fractions of embryonic cells for studying protein shuttling.

机构信息

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Moscow, Russia.

出版信息

STAR Protoc. 2021 Apr 21;2(2):100449. doi: 10.1016/j.xpro.2021.100449. eCollection 2021 Jun 18.

Abstract

This protocol for the separation of nuclear and cytoplasmic fractions of cells of embryos was developed to study changes in the intracellular localization of the Zyxin and Ybx1 proteins, which are capable of changing localization in response to certain stimuli. Western blot analysis allows the quantification of changes in the distribution of these proteins between the cytoplasm and nucleus, whereas the posttranslational modifications specific to each compartment can be identified by changes in electrophoretic mobility. For complete details on the use and execution of this protocol, please refer to Parshina et al. (2020).

摘要

本胚胎细胞核质分离方案旨在研究 Zyxin 和 Ybx1 蛋白在细胞内定位的变化,这两种蛋白能够对外界刺激做出定位改变。Western blot 分析可以定量研究这些蛋白在细胞质和细胞核之间的分布变化,而特定于每个隔室的翻译后修饰可以通过电泳迁移率的变化来识别。如需详细了解本方案的使用和实施,请参考 Parshina 等人(2020 年)的文献。

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