Abstract
Classical two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) allows comparison and quantitation of proteomes by visualization of protein patterns using gel stains and comparative image analysis. The introduction of fluorescent reagents for protein labeling (difference in-gel electrophoresis or DIGE) has brought substantial improvement in this field. It provides multiplexing of up to three samples in one gel, higher sensitivity compared to normal protein staining methods, and a higher linear range for quantitation. This article gives detailed protocols for 2D-DIGE, including both minimal as well as saturation labeling.
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Acknowledgements
This work was supported by the Bundesministerium für Bildung und Forschung (NGFN, FZ 01GS08143) as well as the European Regional Development Fund (ERDF) of the European Union and the Ministerium für Innovation, Wissenschaft und Forschung des Landes Nordrhein-Westfalen (ParkChip, FZ 280381102).
Thanks to Dr. Jan Heise (NH DyeAGNOSTICS GmbH) for providing S-Dyes and G-Dyes.
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May, C., Brosseron, F., Chartowski, P., Meyer, H.E., Marcus, K. (2012). Differential Proteome Analysis Using 2D-DIGE. In: Marcus, K. (eds) Quantitative Methods in Proteomics. Methods in Molecular Biology, vol 893. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-885-6_6
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DOI: https://doi.org/10.1007/978-1-61779-885-6_6
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Publisher Name: Humana Press, Totowa, NJ
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