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 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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Acknowledgment
This study was supported by grants from Spain (BFU-2008-02084/BMC), European Union (FEDER), ISCIII (RETICRD06/0020), and Generalitat de Catalunya (2009SGR1059). MCR, FAP, and IT are supported by a doctoral fellowship from IDIBELL, Generalitat de Catalunya, and 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. (2019). 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) Electrophoretic Separation of Proteins. Methods in Molecular Biology, vol 1855. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8793-1_22
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DOI: https://doi.org/10.1007/978-1-4939-8793-1_22
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