Springer Protocols: Abstract: Isolation and Preparation of Chloroplasts from Arabidopsis thaliana Plants

Isolation and Preparation of Chloroplasts from Arabidopsis thaliana Plants

By: Sybille E Kubis³, Kathryn S Lilley⁴, Paul Jarvis³

Abstract

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A major area of research in the postgenomic era has been the proteomic analysis of various subcellular and suborganellar compartments. The success of these studies is to a large extent dependent upon efficient protocols for the preparation of highly pure organelles or suborganellar components. Here we describe a simple, rapid, and low-cost method for isolating a high yield of Arabidopsis chloroplasts. The method can readily be applied to wild-type plants and different mutants, and at different developmental stages ranging from 10-day-old seedlings to rosette leaves from older plants. The isolated chloroplast fraction is highly pure, with immunologically undetectable contamination from other cellular organelles. Chloroplasts isolated using the method described here have been successfully used for proteomic analysis, as well as in studies on chloroplast protein import and other aspects of chloroplast biology.

Affiliation(s): (3) University of Leicester, Leicester, Great Britain
(4) University of Cambridge, Cambridge, Great Britain

Book Title: 2D PAGE: Sample Preparation and Fractionation

Series: Methods in Molecular Biology | Volume: 425 | Pub. Date: Jan-25-2008 | Page Range: 171-186 | DOI: 10.1007/978-1-60327-210-0_16

Subject: Protein Science

Key Words: Arabidopsis thaliana - chloroplast isolation - chloroplast proteomics - organelle isolation - Percoll gradient - plastids - polytron homogenizer

References

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