Summary
The use of two-dimensional gel electrophoresis for differential analysis in proteomics was revolutionized by the introduction of 2-D fluorescence difference gel electrophoresis (2-D DIGE). This fluorescence-based technique allows the use of multiplexed samples and an internal standard that virtually eliminates gel-to-gel variability, resulting in increased confidence that differences found between samples are due to real induced changes, rather than inherent biological variation or experimental variability. 2-D DIGE has quickly become the “gold standard” for gel-based proteomics for studying tissues, as well as cell culture and bodily fluids such as serum or plasma.
This chapter will describe the basic 2-D DIGE technique using minimal labeling, image acquisition using high-quality fluorescence scanners, and software capable of analyzing the multiplexed images and normalizing the data using the internal standard.
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Acknowledgments
The authors thank Clayton Randall for his assistance preparing the figures, and Phil Beckett and Rita Marouga for critical review of the manuscript.
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Rozanas, C.R., Loyland, S.M. (2008). Capabilities Using 2-D DIGE in Proteomics Research. In: Liu, B.CS., Ehrlich, J.R. (eds) Tissue Proteomics. Methods in Molecular Biology™, vol 441. Humana Press. https://doi.org/10.1007/978-1-60327-047-2_1
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DOI: https://doi.org/10.1007/978-1-60327-047-2_1
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