Isolating intact chloroplasts from small Arabidopsis samples for proteomic studies |
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Authors: | Jeannette Kley Alexander Muck Wilhelm Boland |
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Affiliation: | a Max Planck Institute for Chemical Ecology, 07743 Jena, Germany b Departamento de Ingeniería Genética, Cinvestav-Irapuato, CP 36821, Irapuato, Guanajuato, Mexico |
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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 2-3 g, yielding more than 300 μg 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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Keywords: | Chloroplast Arabidopsis thaliana Proteomics Biotic stress |
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