Abstract
Two-dimensional difference gel electrophoresis (2D DIGE) is a modified form of 2D electrophoresis (2DE) that allows one to compare two or three protein samples simultaneously on the same gel. The proteins in each sample are covalently tagged with different color fluorescent dyes that are designed to have no effect on the relative migration of proteins during electrophoresis. Proteins that are common to the samples appear as “spots” with a fixed ratio of fluorescent signals, whereas proteins that differ between the samples have different fluorescence ratios. With the appropriate imaging system, difference gel electrophoresis (DIGE) is capable of reliably detecting as little as 0.2 fmol of protein, and protein differences down to ±15%, over a ∼20,000-fold protein concentration range. DIGE combined with digital image analysis therefore greatly improves the statistical assessment of proteome variation. Here we describe a protocol for conducting DIGE experiments, which takes 2–3 days to complete.
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Acknowledgments
The development of DIGE would not have been possible without the efforts and dedication of my students and staff (Mustafa Ünlü, Liz Morgan, Chris Lacenere, Surya Viswanathan, Lei Gong, Mamta Puri, Anupam Goyal, and Susan Down).
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Minden, J.S. (2012). Two-Dimensional Difference Gel Electrophoresis. In: Kurien, B., Scofield, R. (eds) Protein Electrophoresis. Methods in Molecular Biology, vol 869. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-821-4_24
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DOI: https://doi.org/10.1007/978-1-61779-821-4_24
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