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DMSO, an Organic Cleanup Solvent for TCA/Acetone-Precipitated Proteins, Improves 2-DE Protein Analysis of Rice Roots

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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–acetone-precipitated pellets. The improved protocol (TCA/acetone–DMSO) 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–DMSO 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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Acknowledgements

This research project was supported by the Fundamental Research Funds for the Central Universities (Y020110055) and the grant from the Education Department of Jiangsu Province (200910).

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Authors and Affiliations

  1. College of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, the People’s Republic of China

    Yufeng Song, Guiping Wang & Zhenguo Shen

  2. College of Agricultural, Henan University of Science and Technology, Luoyang, 471003, the People’s Republic of China

    Hongxiao Zhang

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  1. Yufeng Song
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  2. Hongxiao Zhang
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  3. Guiping Wang
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  4. Zhenguo Shen
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Correspondence to Zhenguo Shen.

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Song, Y., Zhang, H., Wang, G. et al. DMSO, an Organic Cleanup Solvent for TCA/Acetone-Precipitated Proteins, Improves 2-DE Protein Analysis of Rice Roots. Plant Mol Biol Rep 30, 1204–1209 (2012). https://doi.org/10.1007/s11105-012-0442-6

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  • DOI: https://doi.org/10.1007/s11105-012-0442-6

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