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Title of Journal: Plant Cell Rep

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Abbravation: Plant Cell Reports

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Springer Berlin Heidelberg

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DOI

10.1016/0038-1098(87)90913-6

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1432-203X

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An improved immunofluorescence staining method for

Authors: Yiqiong Li Qingqing Sun Yue Feng Xiaomin Liu Hongbo Gao
Publish Date: 2016/07/29
Volume: 35, Issue: 11, Pages: 2285-2293
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Abstract

As an important technical approach immunofluorescence staining is widely used in the subcellular localization study of interested proteins During the study of the functions of chloroplast division proteins immunofluorescence staining was frequently adopted Previously a method has been developed to study the localization of a chloroplast division protein FtsZ However it is laborious and timeconsuming In this study we report a modified immunofluorescence staining method in which protoplasts were isolated from leaf tissues and then fixed for immunofluorescence staining The time of the experiment was significantly reduced to several hours Furthermore we used correction pen in the fixation procedure and a new way to coat the slide which greatly saved the cost of the experiment With the chloroplast division protein ARC6 as an example we can get a good fluorescence signal Moreover the localization of ARC6 in two chloroplast division mutants arc3 and arc5 and three other plant species such as cabbage radish and pea was also successfully analyzed with our new method Overall the immunofluorescence staining method we reported here is very practical and it significantly facilitates the visualization of the subcellular localization of interested proteins in plant cellsThis work was supported by grants from the Science and Technology Innovation Projects of Beijing Forestry University BLX201424 the National Natural Science Foundation of China 31070162 31501090 and the Beijing Natural Science Foundation 5154031

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