DMSO, an Organic Cleanup Solvent for TCA/Acetone-Precipitated Proteins, Improves 2-DE Protein Analysis of Rice Roots |
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Authors: | Yufeng Song Hongxiao Zhang Guiping Wang Zhenguo Shen |
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Institution: | 1. College of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, the People??s Republic of China 2. College of Agricultural, Henan University of Science and Technology, Luoyang, 471003, the People??s Republic of China
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Abstract: | Protein extraction is a critical step in a two-dimensional electrophoresis (2-DE)-based proteomic analysis. Dimethyl sulfoxide (DMSO), a small organic molecule, is widely used as a solvent in biological sciences. In this study, we modified the cleanup step of the commonly used trichloroacetic acid (TCA)/acetone method of protein extraction by using 20?% DMSO/acetone as a solvent to wash TCA?Cacetone-precipitated pellets. The improved protocol (TCA/acetone?CDMSO) was compared with the TCA/acetone and phenol extraction method based on the protein yield, the number of spots, and resolution on 2-DE maps. TCA/acetone?CDMSO washing increased protein yield, and improved the resolution in 2-DE with less background, streaking, and smearing. This method also produced the highest number of protein spots. To our knowledge, the present study is the first to use DMSO as a cleanup solvent for TCA/acetone-precipitated proteins to enhance their quality prior to 2-DE. |
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