Protein Blotting: The Basic Method and its Role in Viral Diagnosis

  • A. J. Herring
  • J. M. Sharp
Part of the Current Topics in Veterinary Medicine and Animal Science book series (CTVM, volume 29)


Protein blotting involves the transfer of a complex mixture of antigens which have been resolved in a polyacrylamide gel on to a nitrocellulose membrane where the various antigenic components are accessible to analysis with antibody. The technique can be used to detect viral antigens or to dissect the humoural response to pathogens. In this paper the basic technique is described and its uses and pitfalls are discussed.


Viral Antigen Chlamydia Trachomatis Mouse Mammary Tumour Virus Electrophoretic Transfer Infected Sheep 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Alwine, J.C., Kemp, D.J. and Stark, G.R. 1977. Method for detection of specific RNAs in agarose gels by transfer to DBM paper and hybridisation with DNA probes. Proc. Natl. Acad. Sci. USA, 74, 5350–5354.PubMedCrossRefGoogle Scholar
  2. Bastardo, J.W., McKimm-Breschkin, J.L., Sonza, S., Mercer, L.D. and Holmes, I.H. 1981. Preparation and characterisation of antisera to electrophoretically purified SA11 virus polypeptides. Infect. Immun., 34, 641–647.PubMedGoogle Scholar
  3. Burnette, W.N. 1981. ‘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, 195–203.PubMedCrossRefGoogle Scholar
  4. Gershoni, J.M. and Palade, G.E. 1983. Protein Blotting: principles and applications. Anal. Biochem., 131, 1–15.PubMedCrossRefGoogle Scholar
  5. Hames, B.D. 1981. In “Gel electrophoresis of proteins” (Ed. B.D. Hames and D. Rickwood). (IRL Press, Oxford and Washington DC).Google Scholar
  6. Hawkes, R., Niday, E. and Gordon, J. 1982. A Dot-immunobinding assay for monoclonal and other antibodies. Anal. Biochem., 119, 142–147.PubMedCrossRefGoogle Scholar
  7. Herring, A.J., Sharp, J.M., Scott, F.M.M. and Angus, K.W. 1983. Further evidence for a retrovirus as the aetiological agent of sheep pulmonary adenomatosis (jaagsiekte). Vet. Microbiol., 8, 237–249.PubMedCrossRefGoogle Scholar
  8. Hunter, W.M. and Greenwood, F.C. 1962. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature, 194, 495–496.PubMedCrossRefGoogle Scholar
  9. Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680–685.PubMedCrossRefGoogle Scholar
  10. Newhall, W.J., Batteiger, B. and Jones, R.G. 1982. Analysis of the human serological response to proteins of Chlamydia trachomatis. Infect. Immun., 38, 1181–1189.PubMedGoogle Scholar
  11. Perk, K., Hod, I. and Nobel, T.A. 1971. Pulmonary adenomatosis of sheep (jaagsiekte). 1. Ultrastructure of the tumour. J. Natl. Cancer Inst., 4, 525–537.Google Scholar
  12. Perk, K., Michalides, R., Spiegelman, S. and Schlom, J. 1974. Biochemical and morphologic evidence for the presence of RNA tumour virus in pulmonary carcinoma of sheep (jaagsiekte). J. Natl. Cancer Inst., 53, 131–135.PubMedGoogle Scholar
  13. Sharp, J.M. and Herring, A.J. 1983. Sheep pulmonary adenomatosis: demonstration of a protein which cross reacts with the major core proteins of Mason-Pfizer monkey virus and mouse mammary tumour virus. J. gen. Virol., 64, 2323–2327.PubMedCrossRefGoogle Scholar
  14. Southern, E.M. 1975. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol., 98, 503–517.PubMedCrossRefGoogle Scholar
  15. Symington, J., Green, M. and Brackmaim, K. 1981. Immunoautoradiographic detection of proteins after electrophoretic transfer from gels to diazopaper: analysis of adenovirus encoded proteins. Proc. Natl. Acad. Sci. USA, 78, 177–181.PubMedCrossRefGoogle Scholar
  16. Tasheva, B. and Dessev, G. 1983. Artifacts in SDS-polyacrylamide gel electrophoresis due to 2-mercaptoethanol. Anal. Biochem., 129, 98–102.PubMedCrossRefGoogle Scholar
  17. Towbin, J., Staehelin, T. and Gordon, J. 1979. Electrophoretic transfer of proteins from Polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA, 76, 4350–4354.PubMedCrossRefGoogle Scholar
  18. Versteegen, R.J. and Oroszlan, S. 1980. Effect of chemical modification and fragmentation on antigenic deteiminants of internal protein p30 and surface protein gp70 of type C retroviruses. J. Virol., 33, 983–992.PubMedGoogle Scholar

Copyright information

© ECSC, EEC, EAEC, Brussels-Luxembourg 1984

Authors and Affiliations

  • A. J. Herring
    • 1
  • J. M. Sharp
    • 1
  1. 1.Moredun Research InstituteEdinburghScotland

Personalised recommendations