Abstract
This chapter describes the basics, applications, and limitations of two-dimensional gel electrophoresis (2DE) and two-dimensional difference gel electrophoresis (2D-DIGE) for multiplex analysis of distinct proteomes. We also propose a basic protocol for 2D-DIGE, technique that allows the analysis of paired protein extracts, which are labeled with fluorescent Cy3 and Cy5 dyes and electrophoresed with a Cy2-labeled standard extract on the same 2DE gels. Scanning the gels at wavelengths specific for each dye allows direct overlay the two different proteomes and the differences in abundance of specific protein spots can be determined.
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Acknowledgments
Dr. Adriano Aquino and Prof. Daniel Martins-de-Souza are funded by FAPESP (São Paulo Research Foundation), grants 15/09159-8 and 13/08711-3.
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Aquino, A., Guest, P.C., Martins-de-Souza, D. (2017). Simultaneous Two-Dimensional Difference Gel Electrophoresis (2D-DIGE) Analysis of Two Distinct Proteomes. In: Guest, P.C. (eds) Multiplex Biomarker Techniques. Methods in Molecular Biology, vol 1546. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-6730-8_17
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DOI: https://doi.org/10.1007/978-1-4939-6730-8_17
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-6729-2
Online ISBN: 978-1-4939-6730-8
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