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Updated: Aug 20, 2025

Preparation of Chloroplast Sub-compartments from Arabidopsis for the Analysis of Protein Localization by Immunoblotting or Proteomics
Published on: October 19, 2018
Chloroplast Envelope Membrane Subfractionation from Arabidopsis and Pea
Annabel Dischinger1, Serena Schwenkert2
1Plant Molecular Biology (Botany), Faculty of Biology, Ludwig-Maximilians-Universität München, Planegg-Martinsried, Germany.
Abstract:
Due to their endosymbiotic origin, chloroplasts harbor several subcompartments and membrane systems. Each of these has a different protein and lipid composition that dynamically changes either naturally during plant development or induced by environmental stimuli. Here, we describe a protocol for chloroplast envelope membrane subfractionation via discontinuous sucrose gradients, which offers the possibility to separate the different plastid subcompartments for several downstream applications. It is a strong tool for protein sublocalization studies as well as for tracking dynamic movement patterns. Furthermore, it can be combined with in vitro import studies of radioactively labeled proteins, which allows sublocalization of putative envelope proteins independent of the availability of specific antisera.
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