Preparation of envelope membrane fractions from arabidopsis chloroplasts for proteomic analysis and other studies - INRAE - Institut national de recherche pour l’agriculture, l’alimentation et l’environnement
Chapitre D'ouvrage Année : 2011

Preparation of envelope membrane fractions from arabidopsis chloroplasts for proteomic analysis and other studies

Résumé

Plastids are semiautonomous organdies restricted to plants and protists. These plastids are surrounded by a double membrane system, or envelope. These envelope membranes contain machineries to import nuclear-encoded proteins, and transporters for ions or metabolites, but are also essential for a range of plastid-specific metabolisms. Targeted semiquantitative proteomic investigations have revealed specific cross-contaminations by other cell or plastid compartments that may occur during chloroplast envelope purification. This article describes procedures developed to recover highly purified envelope fractions starting from Percoll-purified Arabidopsis chloroplasts, gives an overview of possible cross-contaminations, provides some tricks to limit these cross-contaminations, and lists immunological markers and methods that can be used to assess the purity of the envelope fractions.
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Dates et versions

hal-02810049 , version 1 (06-06-2020)

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Citer

Daniel Salvi, Lucas Moyet, Daphné Berny, Myriam Ferro, Jacques Joyard, et al.. Preparation of envelope membrane fractions from arabidopsis chloroplasts for proteomic analysis and other studies. Chloroplast research in arabidopsis: methods and protocols, Vol II, 775, Humana Press, 2011, Methods in Molecular Biology, 978-1-61779-236-6. ⟨10.1007/978-1-61779-237-3_10⟩. ⟨hal-02810049⟩
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