Improvements for Tissue-Chopping-Based Immunofluorescence Staining Method of Chloroplast Proteins
文献类型： 外文期刊
作者： Wang, Lulu 1 ; Chen, Yajuan 3 ; Niu, Di 1 ; Tang, Mingdong 1 ; An, Jinjie 1 ; Xue, Shanshan 1 ; Liu, Xiaomin 1 ; Gao, Hongbo 1 ;
作者机构： 1.Beijing Forestry Univ, Natl Engn Res Ctr Tree Breeding & Ecol Restorat, Beijing 100083, Peoples R China
2.Beijing Forestry Univ, Coll Biol Sci & Technol, Beijing 100083, Peoples R China
3.Beijing Acad Agr & Forestry Sci, Beijing Key Lab Agr Genet Resources & Biotechnol, Inst Biotechnol, Beijing 100097, Peoples R China
关键词： tissue chopping; immunofluorescence staining; chloroplast protein; low temperature
期刊名称：PLANTS-BASEL （ 影响因子：4.5； 五年影响因子：4.8 ）
ISSN：
年卷期： 2023 年 12 卷 4 期
页码：
收录情况： SCI
摘要： 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.
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