4.7 Article

Improvements for Tissue-Chopping-Based Immunofluorescence Staining Method of Chloroplast Proteins

期刊

PLANTS-BASEL
卷 12, 期 4, 页码 -

出版社

MDPI
DOI: 10.3390/plants12040841

关键词

tissue chopping; immunofluorescence staining; chloroplast protein; low temperature

向作者/读者索取更多资源

Immunofluorescence staining is widely used in the subcellular localization study of proteins. A tissue-chopping-based method for immunofluorescence staining of chloroplast proteins is improved by direct observation of stained tissues using a confocal microscope without tissue lysis and by maintaining a low temperature throughout the process. This method has been successfully applied to the staining of FtsZ1 in wildtype and mutant Arabidopsis, as well as in other plants, leading to further improvement of the tissue-chopping-based immunofluorescence staining method.
Immunofluorescence staining is a very common method for the subcellular localization study of proteins. A tissue-chopping-based immunofluorescence staining method for chloroplast proteins overcomes the restriction of plant cell wall, makes the operation simpler, and uses less experimental materials. Here we provide some improvements for this method. We found that the stained tissues can be directly observed with a confocal microscope without tissue lysis. Samples maintained at a low temperature (0-4 degrees C) throughout the process can reduce the intensity of chlorophyll autofluorescence and the background signal. A low temperature is also good for the storage of the sample. Fluorescence signal of the stained samples can be kept for several weeks if they are stored at -20 degrees C. FtsZ is an essential component of the chloroplast division apparatus. We demonstrated this method with the immunofluorescence staining of FtsZ1 in wildtype Arabidopsis and some chloroplast division mutants. We also successfully tested this method by the immunofluorescence staining of FtsZ1 in many other plants, including woody plants. With these procedures, the performance of tissue-chopping-based immunofluorescence staining method are further improved.

作者

我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。

评论

主要评分

4.7
评分不足

次要评分

新颖性
-
重要性
-
科学严谨性
-
评价这篇论文

推荐

暂无数据
暂无数据