Abstract
Plant tissues contain high levels of nonprotein contaminants such as lipids, phenolic compounds, and polysaccharides among others, which interfere with protein extraction and electrophoretic separation. Preparation of good-quality protein extracts is a critical issue for successful electrophoretic analysis. Here, we describe a three-step method for protein extraction from lipid-rich plant tissues, which is suitable for both 1-D and 2-D electrophoresis and is compatible with downstream applications. The protocol includes prefractionation, filtration, and TCA/acetone precipitation steps prior to protein resolubilization.
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Acknowledgements
This work was supported by the Spanish Ministry of Science and Innovation (MICINN) (ERDF-cofinanced projects AGL2008-00517, BFU2011-22779, and PIE-200840I186) and the Junta de Andalucía (ERDF-cofinanced projects P2010-CVI5767 and P2010-AGR6274). A. Zienkiewicz thanks the CSIC for providing JAEdoc grant funding.
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Zienkiewicz, A., Rejón, J.D., de Dios Alché, J., Rodríguez-García, M.I., Castro, A.J. (2014). A Protocol for Protein Extraction from Lipid-Rich Plant Tissues Suitable for Electrophoresis. In: Jorrin-Novo, J., Komatsu, S., Weckwerth, W., Wienkoop, S. (eds) Plant Proteomics. Methods in Molecular Biology, vol 1072. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-631-3_7
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DOI: https://doi.org/10.1007/978-1-62703-631-3_7
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-630-6
Online ISBN: 978-1-62703-631-3
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