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Two-dimensional difference gel electrophoresis
Open accessAbstract
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, DIGE is capable of reliably detecting as little as 0.5 fmol of protein, and protein differences down to ± 15%, over a >10,000-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 d to complete.
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Cited by
Advances in Urinary Proteome Analysis and Biomarker Discovery
Fliser, D., Novak, J., Thongboonkerd, V., Argile[Combining Acute Accent]s, A. G. A., Jankowski, V., Girolami, M. A., Jankowski, J., & Mischak, H. (2007) · Journal of the American Society of Nephrology · doi:10.1681/asn.2006090956 · Cited by 286
Difference gel electrophoresis
Timms, J. F., & Cramer, R. (2008) · PROTEOMICS · doi:10.1002/pmic.200800298 · Cited by 267
Two-dimensional gel electrophoresis in proteomics: A tutorial
Rabilloud, T., & Lelong, C. (2011) · Journal of Proteomics · doi:10.1016/j.jprot.2011.05.040 · Cited by 258
Salanova, M., Gelfi, C., Moriggi, M., Vasso, M., Viganò, A., Minafra, L., Bonifacio, G., Schiffl, G., Gutsmann, M., Felsenberg, D., et al. (2014) · The FASEB Journal · doi:10.1096/fj.14-252825 · Cited by 68
Metadata
- Authors
- Viswanathan, S., Ünlü, M., & Minden, J. S.
- Venue
- Nature Protocols
- Published
- 2006-08
- Volume
- 1
- Issue
- 3
- Pages
- 1351-1358
- ISSN
- 1750-2799 (ISSN-L)
- 1754-2189
- OpenAlex
- W1979542564
- Cited by
- 213
- References
- 0
- FWCI
- 6.81
- Type
- journal article
- Language
- en
- Open access
- green
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