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Analytical Electrophoresis

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Characterization of Proteins

Part of the book series: Biological Methods ((BM))

Abstract

The first true use of electrophoresis for the separation of proteins developed from the pioneering work of Tiselius (1) in 1937, but the concepts and developments that led to the introduction of the technique occurred in the mid-nineteenth century with Faraday’s laws of electrolysis. These laws predated the first accepted demonstration of the chromatographic technique by about 50 years. From Faraday’s laws the concept of transport numbers was developed by Hittorf in 1850. In 1870, Kohlrausch demonstrated the measurement of conductivity using an ac Wheatstone bridge system with a telephone receiver as the null indicating device. From conductivities, the concept of ionic mobilities was developed. These three concepts are fundamental to the development of electrophoresis. The moving boundary method of Tiselius, however, although a superb separation method, was a very difficult technique to control and had the disadvantage that only the slowest and fastest migrating species could be obtained in pure form and then only in small quantities. It was therefore not suited for preparative use. Interestingly, it was Martin’s development of partition and paper chromatography (2,3) that gave the stimulus to develop electrophoretic methods capable of separating components into zones using materials similar to those used in chromatography, Development was rapid, and there is now a powerful armoury of methods suitable for protein separation.

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References

  1. A. Tiselius, Trans. Faraday Soc. 33, 524 (1937).

    Article  CAS  Google Scholar 

  2. A. J. P. Martin and R. L. M. Synge, Biochem. J. 35, 1358 (1941).

    PubMed  CAS  Google Scholar 

  3. R. Consdon, A. H. Gordon, and A. J. P. Martin, Biochem. J. 38, 224 (1944).

    Google Scholar 

  4. I. Smith, ed., Chromatographic and Electrophoretic Techniques vol. 2, Zone Electrophoresis 4th ed., Heinemann, London (1976).

    Google Scholar 

  5. O. Gaal, G. A. Medgyesi, and L. Vereczkey, Electrophoresis in the Separation of Biological Macromolecules Wiley-Interscience, New York (1980).

    Google Scholar 

  6. H. Svensson, Acta. Chem. Scand. 15, 325 (1961).

    Article  CAS  Google Scholar 

  7. H. Svensson, Acta. Chem. Scand. 16, 456 (1962).

    Article  CAS  Google Scholar 

  8. A. Kolin, in Methods in Biochemical Analysis vol. 6 (D. Glick, ed.) Inter-science, New York (1958), p. 259.

    Chapter  Google Scholar 

  9. O. Vesterberg, Acta Chem. Scand. 23, 2653 (1969).

    Article  CAS  Google Scholar 

  10. N. Catsimpoolas, ed., Isoelectric Focussing Academic, New York (1976).

    Google Scholar 

  11. N. Catsimpoolas and J. W. Drysdale, eds., Biological and Biomedical Applications of Isoelectric Focusing Plenum, New York, London (1977).

    Google Scholar 

  12. P. G. Righetti and J. W. Drysdale, in Laboratory Techniques in Biochemistry and Molecular Biology (T. S. Work and E. Work, eds.) Elsevier, Amsterdam (1976).

    Google Scholar 

  13. H. Svensson, Acta Chem. Scand. 15, 325 (1961).

    Article  CAS  Google Scholar 

  14. B. J. Radola, Electrophoresis 1, 43 (1980).

    Article  CAS  Google Scholar 

  15. R. C. Allen, Electrophoresis 1, 32 (1980).

    Article  CAS  Google Scholar 

  16. A. Rosen, K. Ek, and P. Aman, J. Immunol. Meth. 28, (1979).

    Google Scholar 

  17. R. C. Allen, P. M. Oulla, P. Arnaud, and J. S. Baumstark, in Electrofocussing and Isotachophoresis (B. J. Radola and D. Graesslin, eds.) W. de Gruyter, Berlin (1977), pp. 255.

    Google Scholar 

  18. A. Gorg, W. Postel, and R. Westmeier, Anal. Brochem. 89, 60 (1979).

    Article  Google Scholar 

  19. B. J. Radola, in Electrophoresis ’79 (B. J. Radola, ed.) W. de Gruyter, Berlin (1980), p. 79.

    Google Scholar 

  20. B. J. Radola, in Electrophoretic Techniques (C. F. Simpson and M. Whittaker, eds.) Academic, London (1983).

    Google Scholar 

  21. B. Bjelliquist, K. Ek, P. G. Righetti, E. Gianazzi, E. Gory, R. Westermeister, and W. Postel, J. Biochem. Biophys. Methods 6, 317 (1982).

    Article  Google Scholar 

  22. V. Gasparic, B. Bjellqvist, and A. Rosengren, Various patents, see LKB application note 321.

    Google Scholar 

  23. Y. Walbroehl and J. W. Jorgenson, J. Chromatog. 315, 135 (1984)

    Article  CAS  Google Scholar 

  24. J. W. Jorgenson and K. D. Lukacs, Science 222, 266 (1983).

    Article  PubMed  CAS  Google Scholar 

  25. S. Hjerten, J. Chromatog. 270, 1 (1983).

    Article  CAS  Google Scholar 

  26. S. Hjerten, Acta Chem. Scand. B36, 203 (1982).

    CAS  Google Scholar 

  27. M. E. Himmel and P. G. Squire, Int. J. Peptide Protein Res. 17, 365 (1981).

    Article  CAS  Google Scholar 

  28. F. Everaerts and Th. P. E. M. Verheggen, in Electrophoretic Techniques (C. F. Simpson and M. Whittaker, eds.) Academic, London (1983), pp. 149–196.

    Google Scholar 

  29. C. J. Holloway, R. V. Battersby, B. Bussenschutt, G. Bulge, and J. Lustorff, in Analytical and Preparative Isotachophoresis (C. J. Holloway, ed.) Walter de Gruyter, Berlin (1984), pp. 271–289.

    Google Scholar 

  30. J. Bours, in Analytical and Preparative Isotachophoresis (C. J. Holloway, ed.) Walter de Gruyter, Berlin (1984), pp. 315–324.

    Google Scholar 

  31. C. H. Brogren, in Electrofocussing and Isotachophoresis (B. J, Radola and D. Grasslin, eds.) W. de Gruyter, Berlin (1977), pp. 549–558.

    Google Scholar 

  32. O. Smithies and M. D. Poulik, Nature 177, 1033 (1956).

    Article  PubMed  CAS  Google Scholar 

  33. G. Dale and A. L. Latner, Clin. Chem. Acta 24, 61 (1969).

    Article  CAS  Google Scholar 

  34. P. H. O’Farrell, J. Biol. Chem. 250, 4007 (1975).

    PubMed  Google Scholar 

  35. N. G. Anderson and N. L. Anderson, Anal. Biochem. 85, 331 (1978) [see also M. J. Dunn and A. H. M. Burghes, Electrophoresis 4, 973 1983, for a recent review with 215 references].

    Article  PubMed  CAS  Google Scholar 

  36. P. Z. O’Farrell, H. M. Goodman, and P. H. O’Farrell, Cell 12, 1133 (1977).

    Article  PubMed  Google Scholar 

  37. C. Wrigley, Biochem. Genetics 4, 509 (1970).

    Article  CAS  Google Scholar 

  38. K. G. Kenrick and J. Margolis, Anal. Blochem. 33, 204 (1970).

    Article  CAS  Google Scholar 

  39. D. H. Leaback, in Isoelectric Focussing (J. P. Arbuthnott and J A. Beeley, eds.) Butterworths, London (1975), pp. 201.

    Google Scholar 

  40. H. Stegmann, H. Francksen, and V. Macko, Z. Nuturforsch 28, 722 (1973).

    Google Scholar 

  41. T. Barrett and H. J. Gould, Eur. J. Biochem. 93, 329 (1979).

    Google Scholar 

  42. G. P. Tuszynski, C. A Buck, and L. Warren, Anal. Biochem. 93, 329 (1979).

    Article  PubMed  CAS  Google Scholar 

  43. C. H. Brogren, in Electrophoresis and Isotachophoresis (B J. Radola and D. Grasslin, ed.) W de Gruyter, Berlin (1977), pp. 549.

    Google Scholar 

  44. P. D. Eckersall and J. A. Beeley, Electrophoresis 1, 62 (1980).

    Article  CAS  Google Scholar 

  45. J. S. Fawcett, in Electrophoretic Techniques (C. F. Simpson and M. Whittaker, eds.) Academic, London (1983), pp. 57–80.

    Google Scholar 

  46. A. Gorg, W. Postel, and R. Westermeier, in Electrophoresis ’79 (B. J. Radola, ed.) W. de Gruyter, Berlin (1980), p. 67.

    Google Scholar 

  47. P. E. Righetti and F Chellemi, J. Chromatog. 157, 253 (1978).

    Article  Google Scholar 

  48. T. Bibring and J. Baxendale, Anal. Biochem. 85, 1 (1978).

    Article  PubMed  CAS  Google Scholar 

  49. R. E. Allen, K. C. Marsak, and P. K. McAllister, Anal. Biochem. 104, 494 (1980).

    Article  PubMed  CAS  Google Scholar 

  50. H. D. Kay, Electrophoresis 2, 304 (1981)

    Article  Google Scholar 

  51. S. Peats, Analyt. Biochem. 140, 178 (1984).

    Article  PubMed  CAS  Google Scholar 

  52. T. Marshall, Analyt. Biochem. 136, 340 (1984).

    Article  PubMed  CAS  Google Scholar 

  53. F. Lasne, O. Benzerra, and Y. Lasne, Analyt. Biochem. 132, 338 (1983).

    Article  PubMed  CAS  Google Scholar 

  54. J. Bossinger, M. J. Miller, K. P. Vo, P. Geudushik, and N. H. Xung, J. Biol. chem. 254, 7960 (1975).

    Google Scholar 

  55. E. Kitatoe, M. Mayhara, N. Kiraska, H. Ueta, and K. Utumsi, Anal. Biochem. 134, 295 (1984).

    Article  Google Scholar 

  56. J. D. Weinstein, T: G. Obrig, and J. B. Weiggle, J. Chromatog. 157, 198 (1978).

    Google Scholar 

  57. E. M. Southern, J. Mol. Biol. 98, 503 (1975).

    Article  PubMed  CAS  Google Scholar 

  58. J. Renart, J. Reiser, and G. R. Stark, Proc. Natl. Acad. Sci. USA 76, 4350 (1979).

    Article  Google Scholar 

  59. H. Towbin, T Staehelin, and J. Gordon, Proc. Natl. Acad. Sci. USA 76, 4350 (1979).

    Article  PubMed  CAS  Google Scholar 

  60. J. M. Gershoni and G. E. Palade, Anal. Biochem. 131, 1 (1983).

    Article  PubMed  CAS  Google Scholar 

  61. J. Symington, in Two-Dimensional Gel Electrophoresis of Proteins: Methods and Applications (J. E. Celis and R. Bravo, eds.) Academic, New York (1983).

    Google Scholar 

  62. H.H. Laver and D. McManigill, Anal. Chem. 58, 166 (1986).

    Article  Google Scholar 

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Author information

Authors and Affiliations

  1. Department of Chemistry, Chelsea College, Universzty of London, London, UK

    Colin Simpson

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  1. Colin Simpson
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Editors and Affiliations

  1. Pafra Ltd., Cambridge, UK

    Felix Franks

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© 1988 Humana Press Inc.

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Simpson, C. (1988). Analytical Electrophoresis. In: Franks, F. (eds) Characterization of Proteins. Biological Methods. Humana Press. https://doi.org/10.1007/978-1-59259-437-5_12

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  • DOI: https://doi.org/10.1007/978-1-59259-437-5_12

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-109-8

  • Online ISBN: 978-1-59259-437-5

  • eBook Packages: Springer Book Archive

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