期刊
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY
卷 52, 期 5, 页码 701-704出版社
SAGE PUBLICATIONS LTD
DOI: 10.1177/002215540405200514
关键词
carboxyfluorescein diacetate; intact organelles; Arabidopsis; chloroplast; glucosinolates; protein import
类别
We developed a rapid and reliable technique for specifically staining intact chloroplasts using the fluorescent dye carboxyfluorescein diacetate. Intact, import-competent chloroplasts were isolated simply and rapidly from soil-grown Arabidopsis thaliana plants, with yields of 20 +/- 5 mug chlorophyll per g FW, greater than previously reported yields from soil-grown Arabidopsis. Traditional chloroplast isolation buffers sometimes contain low concentrations (<10 mM) sodium ascorbate as a general-purpose anti-oxidant, but we found that only Arabidopsis chloroplasts isolated in the presence of high concentrations (50-100 mM) of sodium ascorbate in the initial grinding buffer were import-competent.
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