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  • Title: Isolating intact chloroplasts from small Arabidopsis samples for proteomic studies.
    Author: Kley J, Heil M, Muck A, Svatos A, Boland W.
    Journal: Anal Biochem; 2010 Mar 15; 398(2):198-202. PubMed ID: 19917261.
    Abstract:
    We have established a method for the isolation of chloroplasts from Arabidopsis thaliana that allows proteomic studies in the context of biotic stress with small amounts of starting material. Employing a 50% Percoll layer to separate crude filtrates, the required leaf material was reduced to 2-3g, yielding more than 300 microg of chloroplast proteins. The quality of this fraction was confirmed by immunological, enzymatic, and gel-based assays. This protocol provides intact chloroplasts from Arabidopsis plants with a high degree of integrity and purity as well as sufficient protein recovery, thereby enabling studies of plant-herbivore or plant-pathogen interactions.
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