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从拟南芥中免疫纯化线粒体。

Immunopurification of Mitochondria from Arabidopsis.

机构信息

State Key Laboratory of Plant Genomics and National Center for Plant Gene Research, Institute of Genetics and Developmental Biology, The Innovative Academy of Seed Design, Chinese Academy of Sciences, Beijing, China.

University of Chinese Academy of Sciences, Beijing, China.

出版信息

Curr Protoc. 2021 Feb;1(2):e34. doi: 10.1002/cpz1.34.

Abstract

Isolation of mitochondria from plant tissues, especially green leaves, is particularly difficult because of chloroplast contamination. Compared to other techniques for purifying plant mitochondria, the immunopurification method has a number of advantages: it rapidly purifies the mitochondria within several minutes, avoids loss of mitochondrial metabolites, eliminates most other organelles (especially the chloroplasts), keeps the isolated mitochondria intact, requires little starting material, and has the potential to purify tissue-specific mitochondria. Here, we describe a rapid immunopurification protocol for mitochondrial isolation that employs a protein specifically targeted to the outer mitochondrial membrane that acts as an immunopurification handle. © 2021 Wiley Periodicals LLC.

摘要

从植物组织中分离线粒体,特别是从绿色叶片中分离,由于叶绿体的污染,特别困难。与其他纯化植物线粒体的技术相比,免疫纯化方法具有许多优点:它可以在几分钟内快速纯化线粒体,避免线粒体代谢物的损失,消除大多数其他细胞器(特别是叶绿体),保持分离的线粒体完整,所需起始材料少,并且具有纯化组织特异性线粒体的潜力。在这里,我们描述了一种快速的免疫纯化线粒体分离方法,该方法使用一种专门针对外膜的蛋白质作为免疫纯化的把手。© 2021Wiley Periodicals LLC.

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