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2-D Protein Gel Electrophoresis

An Overview

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2-D Proteome Analysis Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 112))

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Abstract

Two-dimensional electrophoresis (2-D) of proteins used to be an art practiced by a few researchers, and their worldwide meetings could be held in a side room of a medium-sized hotel. With the rapidly growing volume of sequence data produced by the genome projects and the development of new mass spectrometry methods, high-resolution protein analysis has become an important tool in molecular biology. High-resolution 2-D can reveal virtually all proteins present in a cell or tissue at any given time, including those with posttranslational modifications and rapid turnover rates. With the new analytical tools and the genomic and protein database networks, large-scale studies can be performed on the actual gene products or proteins, in their precursor, mature, and modified forms. This task has lately been called the proteome project. The proteome projects are the necessary complement to genome analysis, and aim to identify and characterize all proteins expressed by an organism or a tissue (1).

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References

  1. Wilkins, M. R., Sanchez, J. C., Gooley, A. A., Appel, R. D., Humphrey-Smith, E., Hochstrasser, D. S., and Williams, K. L. (1995) Progress with proteome projects: why all proteins expressed by a genome should be identified and how to do it. Biotechnol. Gene. Eng. Rev. 13, 19–50.

    Google Scholar 

  2. Klose, J. and Kobalz, U. (1995) Two-dimensional electrophoresis of proteins: an updated protocol and implications for functional analysis of the genome. Electrophoresis 16, 1034–1059.

    Article  PubMed  CAS  Google Scholar 

  3. Celis, J. E., Rasmussen, H. H., Gromov, P., Olsen, E., Madsen, P., Leffers, H., Honor, B., Dejgaard, K., Vorum, H., Kristensen, D. B., Ostergaard, M., Haunse, A., Jensen, N. A., Celis, A., Basse, B., Lauridsen, J. B., Ratz, G. P., Anderson, A. H., Walbum, E., Kjaergaard, I., Andersen, I., Puype, M., Van Damme, J., and Vanderkerckhove, J. (1995) The human keratinocyte two-dimensional gel protein database (update 1995): Mapping components of signal transduction pathways. Electrophoresis 16, 2177–2240.

    Article  PubMed  CAS  Google Scholar 

  4. Pasquali, C., Frutiger, S., Wilkins, M. R., Hughes, G. J., Appel, R. D., Bairoch, A., Schaller, D., Sanchez, J.-C., and Hochstrasser, D. F. (1996) Two-dimensional gel electrophoresis of Escherichia coli homogenates: the Escherchia coli SWISS-2DPAGE database. Electrophoresis 17, 547–555.

    Article  PubMed  CAS  Google Scholar 

  5. Macko, V. and Stegemann, H. (1969) Mapping of potato proteins by combined electrofocusing and electrophoresis. Identification of varieties. Hoppe-Seylerís Z. Physiol. Chem. 350, 917–919.

    CAS  Google Scholar 

  6. Klose, J. (1975) Protein mapping by combined isoelectric focusing and electrophoresis of mouse tissues. Humangenetik 26, 231–243.

    PubMed  CAS  Google Scholar 

  7. O’Farrell, P. H. (1975) High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. 250, 4007–4021.

    Google Scholar 

  8. Anderson, N. G. and Anderson, N. L. (1978) Analytical techniques for cell fractions XXI. Two-dimensional analysis of serum and tissue proteins: Multiple isoelectric focusing. Anal. Biochem. 85, 331–340.

    Article  PubMed  CAS  Google Scholar 

  9. Anderson, N. L. and Anderson, N. G. (1978) Analytical techniques for cell Fractions XXII. Two-dimensional analysis of serum and tissue proteins: Multiple gradientslab gel electrophoresis. Anal. Biochem. 85, 341–354.

    Article  PubMed  CAS  Google Scholar 

  10. Bjellqvist, B., Ek, K., Righetti, P. G., Gianazza, E., Görg, A., Westermeier R., et al. (1982) Isoelectric focusing in immobilized pH gradients: principle, methodology and some applications. J. Biochem. Biophys. Methods 6, 317–339.

    Article  PubMed  CAS  Google Scholar 

  11. Görg, A., Postel, W., and Günther, S. (1988) The current state of two-dimensional electrophoresis with immobilized pH gradients. Electrophoresis 9, 531–546.

    Article  PubMed  Google Scholar 

  12. Blomberg, A., Blomberg, L., Norbeck, J., Fey, S. J., Larsen, P. M., Roepstorff, P., Degand, H., Boutry, M., Posch, A., and Görg, A. (1995) Interlaboratory reproducibility of yeast protein patterns analysed by immobilized pH gradient two-dimensional gel electrophoresis. Electrophoresis 16, 1935–1945.

    Article  PubMed  CAS  Google Scholar 

  13. Rabilloud, T., Valette, C., and Lawrence, J. J. (1994) Sample application by in-gel rehydration improves the resolution of two-dimensional electrophoresis with immobilized pH gradients in the first dimension. Electrophoresis 15, 1552–1558.

    Article  PubMed  CAS  Google Scholar 

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© 1999 Humana Press Inc., Totowa, NJ

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Fichmann, J., Westermeier, R. (1999). 2-D Protein Gel Electrophoresis. In: Link, A.J. (eds) 2-D Proteome Analysis Protocols. Methods in Molecular Biology, vol 112. Humana Press. https://doi.org/10.1385/1-59259-584-7:1

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  • DOI: https://doi.org/10.1385/1-59259-584-7:1

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-524-9

  • Online ISBN: 978-1-59259-584-6

  • eBook Packages: Springer Protocols

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