Abstract
Two-dimensional electrophoresis (2-DE) is a classical separation technique for complex protein mixtures based on a first-separation step by isoelectric point (pI) followed by SDS-PAGE. Due to its high resolving power (including protein isoforms) and the capability to display post-translational modifications, 2-DE has been the technique of choice in many studies focused in characterizing allergens. In this chapter, we describe a standard protocol which enables efficient protein separation and subsequent protein identification by tandem mass spectrometry (MALDI-MS/MS).
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Acknowledgments
This work was supported by grants from the Instituto de Salud Carlos III (FIS PI070537, IF08/3667-1, PI11-02239, PI14/01917, PI11/01401, PI11/02432, PI13/01873, PI13/01746, PI13/01581, PI14/01650, PI14/01841) by PT13/0001/0013, IDCSalud (3371/002) and Redes Temáticas de Investigación Cooperativa (FONDOS FEDER, RD06/0014/1015, RD12/0042/0071).
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de la Cuesta, F., Alvarez-Llamas, G., Barderas, M.G. (2017). Two-Dimensional Electrophoresis and Identification by Mass Spectrometry. In: Lin, J., Alcocer, M. (eds) Food Allergens. Methods in Molecular Biology, vol 1592. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6925-8_6
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DOI: https://doi.org/10.1007/978-1-4939-6925-8_6
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