Abstract
Polyacrylamide gel electrophoresis (PAGE) is one of the most powerful tools used for protein analysis. We describe the use of Tris–acetate buffer and 3–15% polyacrylamide gradient gels to simultaneously separate proteins in the mass range of 10–500 kDa. We show that this system is highly sensitive, it has good resolution and high reproducibility, and that it can be used for general applications of PAGE such as Coomassie Brilliant Blue staining and immunoblotting. Moreover, we describe how to generate mini Tris–acetate polyacrylamide gels to use them in miniprotein electrophoresis systems. These economical gels are easy to generate and to manipulate and allow a rapid analysis of proteins. All these features make the Tris–acetate–PAGE system a very helpful tool for protein analysis.
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Acknowledgments
This study was supported by grants from Spain (BFU2008-02084 and BFU2011-22498), European Union (FEDER), ISCIII (RETICRD06/0020), and Generalitat de Catalunya (2009SGR1059). M.C.R, F. A. P., and I. T. are supported by a doctoral fellowship from IDIBELL, Generalitat de Catalunya, and by the Juan de la Cierva program, respectively.
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Cubillos-Rojas, M., Amair-Pinedo, F., Tato, I., Bartrons, R., Ventura, F., Rosa, J.L. (2012). Tris–Acetate Polyacrylamide Gradient Gels for the Simultaneous Electrophoretic Analysis of Proteins of Very High and Low Molecular Mass. In: Kurien, B., Scofield, R. (eds) Protein Electrophoresis. Methods in Molecular Biology, vol 869. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-821-4_17
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DOI: https://doi.org/10.1007/978-1-61779-821-4_17
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Online ISBN: 978-1-61779-821-4
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