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Techniques in Molecular Biology pp 49–61Cite as

Protein Blotting

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Abstract

Polyacrylamide gel electrophoresis has proved to be an extremely powerful tool for the analysis of complex mixtures of proteins (see Chapter 2). However, while the resolving power of this method has progressively increased (the two-dimensional gel system of O’Farrell1 for example being capable of resolving more than 1600 different proteins) any further analysis of the separated proteins has been limited. The majority of the protein molecules lie buried within the gel matrix and are therefore not readily accessible to further investigation. Though methods have been developed to overcome this problem (for example by elution of the proteins2 or by direct in-situ analysis using antisera3,4 or other protein probes5) these methods are in general time-consuming, insensitive and also frequently lead to a loss of resolution.

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Further Reading

  • Schleicher and Schuell Inc., USA. Methods for the transfer of DNA, RNA and protein to nitrocellulose and diazotized paper solid supports. By: Barinaga. M. et al., 1981.

    Google Scholar 

  • Bio-Rad Laboratories.’ southern, Western and Electroblotting’, Bulletin 1080 EG, (June 1981).

    Google Scholar 

References

  1. P.H. O’Farrell and P.Z. O’Farrell, ‘Two dimensional polyacrylamide gel electrophoretic fractionation’ in G. Stein. J. Stein and L.J. Kleinsmith, Chromatin and chromosomal protein research, Methods in Cell Biology 16. (Academic Press, New York, London, Toronto, Sydney. San Francisco, 1977), Part 1, 407–420.

    Google Scholar 

  2. M. Bustin, R.B. Hopkins and I. Isenberg, ‘Immunological relatedness of high mobility group chromosomal proteins from calf thymus’, J. Biol. Chem., 253 (1978), 1694–1699.

    PubMed  CAS  Google Scholar 

  3. W.S. Adari, D. Jurivich and U.W. Goodenough, ‘Localization of cellular antigens in sodium dodecylsulphate-polyacrylamide gels’, J. Cell Biol., 79 (1978), 281–285.

    Article  Google Scholar 

  4. K. Burridge, ‘Changes in cellular glycoproteins after transformation: Identification of specific glycoproteins and antigens in sodium dodecyl sulphate gels’, Proc. Natl. Acad. Sci. USA, 73 (1976), 4457–4461.

    Article  PubMed  CAS  Google Scholar 

  5. M.C. Snabes, A.E. Boyd III and J. Bryan, ‘Detection of actin-binding proteins in human platelets by 125I actin overlay of polyacrylamide gels’, J. Cell Biol., 90 (1981), 809–812.

    Article  PubMed  CAS  Google Scholar 

  6. E.M. Southern, ‘Detection of specific sequences among DNA fragments separated by gel electrophoresis’, J. Mol. Biol., 98 (1975), 503–517.

    Article  PubMed  CAS  Google Scholar 

  7. Ibid.

    Google Scholar 

  8. B. Bowen. J. Steinberg. U.K. Laemmli and H. Weintraub, ‘The detection of DNA-binding proteins by protein blotting’, Nucleic Acids Res., 8 (1980), 1–20.

    Article  PubMed  CAS  Google Scholar 

  9. H. Towbin, T. Stahelin and J. Gordon, ‘Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets. Procedure and some applications’, Proc. Natl. Acad. Sci. USA. 76 (1979), 4350–4354.

    Article  PubMed  CAS  Google Scholar 

  10. Ibid.

    Google Scholar 

  11. W.N. Burnett, ‘“Western blotting”. Electrophoretic transfer of proteins from SDS-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A’, Anal. Biochem., 112 (1981), 195–203.

    Article  Google Scholar 

  12. W. Gibson, ‘Protease-facilitated transfer of high-molecular-weight proteins during electrotransfer to nitrocellulose’, Anal. Biochem., 118 (1981), 1–3.

    Article  PubMed  CAS  Google Scholar 

  13. D.W. Cleveland, S.G. Fischer, M.W. Kirschner and U.K. Laemmli, ‘Peptide mapping by limited proteolysis in sodium dodecyl sulphate and analysis by gel electrophoresis’, J. Biol. Chem., 252 (1977), 1102–1106.

    PubMed  CAS  Google Scholar 

  14. Bowen, Steinberg, Laemmli and Weintraub, ‘The detection of DNA-binding proteins’.

    Google Scholar 

  15. M. Bittner, P. Kupferer and C.F. Morris, ‘Electrophoretic transfer of proteins and nucleic acids from slab gels to diazobenzyloxymethyl cellulose or nitrocellulose sheets’, Anal. Biochem., 102 (1980), 459–471.

    Article  PubMed  CAS  Google Scholar 

  16. T.P. St John and R.W. Davis, ‘Isolation of galactose-inducible DNA sequences from Saccharomyces cerevisiae by differential plaque filter hybridisation’, Cell, 16 (1979), 443–452.

    Article  Google Scholar 

  17. Burnette, “Western blotting”.

    Google Scholar 

  18. J.C. Alwine, D.J. Kemp and G.R. Stark, ‘Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl paper and hybridization with DNA probes’, Proc. Natl Acad. Sci. USA, 74 (1977), 5350–5354.

    Article  PubMed  CAS  Google Scholar 

  19. J. Renart, J. Reiser and G.R. Stark, ‘Transfer of proteins from gels to diazobenzyloxymethyl paper and detection with antisera: A method for studying antibody specificity and antigen structure’, Proc. Natl. Acad. Sci. USA, 76 (1979), 3116–3120.

    Article  PubMed  CAS  Google Scholar 

  20. J. Symington, M. Green and K. Brackman, ‘Immunoautoradiographic detection of proteins after electrophoretic transfer from gels to diazo-paper: Analysis of adenovirus encoded proteins’, Proc. Natl. Acad. Sci. USA, 78 (1981), 177–181.

    Article  PubMed  CAS  Google Scholar 

  21. L. Clarke, R. Hitzeman and J. Carbon, ‘Selection of specific clones from colony banks by screening with radioactive antibody’, Methods in Enzymology, (Academic Press, New York, London), 68 (1979), 436–442.

    Google Scholar 

  22. B.S. Bhullar, J. Hewitt and E.P.M. Candido, ‘The large high mobility group proteins of rainbow trout are localized predominantly in the nucleus and nucleoli of a cultured trout cell line’, J. Biol. Chem., 256 (1981). 8801–8806.

    PubMed  CAS  Google Scholar 

  23. Bowen, Steinberg, Laemmli and Weintraub, ‘The detection of DNA-binding proteins’.

    Google Scholar 

  24. Towbin, Stahelin and Gordon, ‘Electrophoretic transfer of proteins’.

    Google Scholar 

  25. Ibid.

    Google Scholar 

  26. Burnette, “Western blotting”.

    Google Scholar 

  27. Bittner, Kupferer and Morris, ‘Electrophoretic transfer of proteins’.

    Google Scholar 

  28. D. Denharft, ‘A membrane-filter technique for the detection of complementary DNA’, Biochem. Biophys. Res. Commun., 23 (1966), 641–646.

    Article  Google Scholar 

  29. Towbin, Stahelin and Gordon, ‘Electrophoretic transfer of proteins’.

    Google Scholar 

  30. Burnette, “Western blotting”.

    Google Scholar 

  31. V. Shen, T.C. King, V. Kumar and B. Daugherty, ‘Monoclonal antibodies to Escherichia coli 50S ribosomes’, Nucleic Acids Res., 8 (1980), 4639–4649.

    Article  PubMed  CAS  Google Scholar 

  32. Bowen, Steinberg, Laemmli and Weintraub, ‘The detection of DNA-binding proteins’.

    Google Scholar 

  33. R.S. Gilmour, A. Lang, J.R. Gu, C. Johnston and J. Paul, DNA binding proteins in chromatin structure and function’, Cell Biol. Int. Repts., 5 (1981), 45–53.

    Article  CAS  Google Scholar 

  34. Bowen, Steinberg. Laemmli and Weintraub, ‘The detection of DNA-binding proteins’.

    Google Scholar 

  35. Gilmour, Lang, Gu, Johnston and Paul, ‘DNA-binding proteins’.

    Google Scholar 

  36. Burnette, “Western blotting”.

    Google Scholar 

  37. Bowen, Steinberg, Laemmli and Weintraub, ‘The detection of DNA-binding proteins’.

    Google Scholar 

  38. H.A. Erlich, J.R. Levinson, S.N. Cohen and H.O. McDevitt, ‘Filter affinity transfer. A new technique for the in-situ identification of proteins in gels’, J. Biol. Chem., 254 (1979), 12240–12247.

    PubMed  CAS  Google Scholar 

  39. Burnette, “Western blotting”.

    Google Scholar 

  40. Bowen, Steinberg, Laemmli and Weintraub, ‘The detection of DNA-binding proteins’.

    Google Scholar 

  41. Erlich, Levinson, Cohen and McDevitt, ‘Filter affinity transfer’.

    Google Scholar 

  42. Towbin, Stahelin and Gordon, ‘Electrophoretic transfer of proteins’.

    Google Scholar 

  43. Burnette, ‘“Western blotting”’.

    Google Scholar 

  44. Symington, Green and Brackman, ‘Immunoautoradiographic detection of proteins’.

    Google Scholar 

  45. Bhullar, Hewitt and Candido, ‘The large high mobility group proteins’.

    Google Scholar 

  46. Erlich, Levinson, Cohen and McDevitt, ‘Filter affinity transfer’.

    Google Scholar 

  47. Burnette, ‘“Western blotting”’.

    Google Scholar 

  48. Renart, Reiser and Stark, ‘Transfer of proteins’.

    Google Scholar 

  49. Bhullar, Hewitt and Candido, ‘The large high mobility group of proteins’.

    Google Scholar 

  50. Towbin, Stahelin and Gordon, ‘Electrophoretic transfer of proteins’.

    Google Scholar 

  51. Erlich, Levinson, Cohen and McDevitt, ‘Filter affinity transfer’.

    Google Scholar 

  52. Towbin, Stahelin and Gordon, ‘Electrophoretic transfer of proteins’.

    Google Scholar 

  53. Ibid.

    Google Scholar 

  54. R.T.M.J. Vaessen, J. Kreike and G.S.P. Groot, ‘Protein transfer to nitrocellulose filters’, FEBS Letts., 124 (1981), 193–196.

    Article  CAS  Google Scholar 

  55. Bowen, Steinberg, Laemmli and Weintraub, ‘The detection of DNA-binding proteins’.

    Google Scholar 

  56. Gilmour, Lang, Gu, Johnston and Paul, ‘DNA-binding proteins’.

    Google Scholar 

  57. Bowen, Steinberg, Laemmli and Weintraub, ‘The detection of DNA-binding proteins’.

    Google Scholar 

  58. Ibid.

    Google Scholar 

  59. Erlich, Levinson, Cohen and McDevitt, ‘Filter affinity transfer’.

    Google Scholar 

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Authors
  1. Keith Gooderham
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Editors and Affiliations

  1. Department of Biological Sciences, The Hatfield Polytechnic, PO Box 109, Hatfield, Herts, AL10 9AB, England

    John M. Walker

  2. Department of Applied Microbial Genetics, Technical University of Denmark, Building 221, 2800, Lyngby, Denmark

    Wim Gaastra

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© 1983 John M. Walker and Wim Gaastra

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Gooderham, K. (1983). Protein Blotting. In: Walker, J.M., Gaastra, W. (eds) Techniques in Molecular Biology. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-6563-1_3

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  • DOI: https://doi.org/10.1007/978-94-011-6563-1_3

  • Publisher Name: Springer, Dordrecht

  • Print ISBN: 978-0-7099-2755-6

  • Online ISBN: 978-94-011-6563-1

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