• Kari Johansen
  • Lennart Svensson
Part of the Springer Protocols Handbooks book series (SPH)


Immunoprecipitation allows the investigator to detect and quantitate antigens in a mixture of proteins or characterize a specific antibody response to already well-characterized proteins. Addition of antibodies to proteins, usually radiolabeled, allows formation of antigen-antibody complexes. After separation from contaminating proteins, the complexes are disassociated and the proteins of interest are separated by sodium dodecyl sulphate -polyacrylamide gel electrophoresis (SDS-PAGE). Size and quantity of proteins may then be analyzed either by autoradiography or a gel scanning procedure. Immunoprecipitation is extremely sensitive and may detect very small amounts of radiolabeled protein (detection level ∼100 pg protein or 100 cpm/protein). Unlabeled proteins may be used if other sensitive detection methods are utilized, e.g., enzymatic activity assays or Western blotting. The advantage of the immunoprecipitation technique vs immunoblotting is the possibility to analyze the immune response to proteins expressed in their native conformation. Radioimmunoprecipitation assay (RIPA) is used routinely for the detection of viral proteins, characterization of monoclonal and polyclonal antibody preparations, and determination of the specificity of the immune response to various pathogens (1-3).


Sodium Salicylate Bromphenol Blue Radiolabeled Amino Acid Unlabeled Protein Immunoprecipitated Protein 
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.


  1. 1.
    Sambrook, J., Fritsch E. F., and Maniatis, T. (eds). (1989) Molecular Cloning: A Laboratory Manual (2nd ed.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.Google Scholar
  2. 2.
    Harlow, E. and Lane, D. (1988) Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.Google Scholar
  3. 3.
    Burleson, F. G., Chambers, T. M., and Wiedbrauk, D. L. (eds). (1992) Virology: A Laboratory Manual. Academic, San Diego, CA.Google Scholar
  4. 4.
    Barrett, A. J. and Salvesen, G. (eds). (1986) Proteinase Inhibitors. Research Monographs in Cell and Tissue Physiology, vol. 12. Elsevier, Amsterdam, The Netherlands.Google Scholar
  5. 5.
    James, G. T. (1978) Inactivation of the protease inhibitor phenylmethylsulfonyl fluoride in buffers. Analyt. Biochem. 86, 574–579.PubMedCrossRefGoogle Scholar
  6. 6.
    Hammond, C. and Helenius, A. (1994) Quality control in the secretory pathway of a misfolded viral membrane glycoprotein involves cycling between the ER, intermediate compartment, and Golgi apparatus. J. Cell Biol. 126, 41–52.PubMedCrossRefGoogle Scholar
  7. 7.
    Hjelm, H., Hjelm, K., and Sj/:oquist, J. (1972) Protein A from Staphylococcus aureus. Its isolation by affinity chromatography and its use as an immunosorbent for isolation of immunoglobulins. FEBS Lett. 28, 73–76.PubMedCrossRefGoogle Scholar
  8. 8.
    Sj/:odahl, J. (1977) Structural studies on the four repetitive Fc-binding regions in protein A from Staphylococcus aureus. Eur. J. Biochem. 78, 471–490.CrossRefGoogle Scholar
  9. 9.
    Goudswaard, J., van der Donk, J. A., Noordzij, A., van Dam, R. H., and Vaerman, J.-P. (1978) Protein A reactivity of various mammalian immunoglobulins. Scand. J. Immunol. 8, 21–28.PubMedCrossRefGoogle Scholar
  10. 10.
    Bj/:orck, L. and Kronvall, G. (1984) Purification and some properties of streptococcal protein G: a novel IgG-binding reagent. J. Immunol. 133, 969–974.Google Scholar
  11. 11.
    Åkerstr/:om, B., Brodin, T., Reis, K., and Björck, L. (1985) Protein G: a powerful tool for binding and detection of monoclonal and polyclonal antibodies. J. Immunol. 135, 2589–2592.Google Scholar
  12. 12.
    Fahnestock, S. R., Alexander, P., Nagle, J., and Filpula, D. (1986) Gene for an immunoglobulin-binding protein from a group G streptococcus. J. Bacteriol. 167(3), 870–880.PubMedGoogle Scholar
  13. 13.
    Roque-Barreira, M. C. and Campos-Neto, A. (1985) Jacalin: an IgA-binding lectin. J. Immunol. 134, 1740–1743.PubMedGoogle Scholar
  14. 14.
    Johansen, K., Granqvist, L., Karlen, K., Stintzing, G., Uhnoo, I., and Svensson, L. (1994) Serum IgA immune response to individual rotavirus polypeptides in young children with rotavirus infection. Arch. Virol. 138, 247–259.PubMedCrossRefGoogle Scholar
  15. 15.
    Studier, F. W. (1973) Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J. Mol. Biol. 79, 237–248.PubMedCrossRefGoogle Scholar
  16. 16.
    Hames, B. D. and Rickwood, D. (eds.) (1981) Gel Electrophoresis of Proteins: A Practical Approach. IRL, Oxford, UK.Google Scholar
  17. 17.
    Ornstein, L. (1964) Disc electrophoresis-I. Background and theory. Ann. NY Acad. Sci. 121, 321–349.PubMedCrossRefGoogle Scholar
  18. 18.
    Davis, B. J. (1964) Disc-electrophoresis II. Method and application to human serum proteins. Ann. NY Acad. Sci. 121, 404–427.PubMedCrossRefGoogle Scholar
  19. 19.
    Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2002

Authors and Affiliations

  • Kari Johansen
    • 1
  • Lennart Svensson
    • 1
  1. 1.Department of VirologySwedish Institute For Infectious Disease ControlSweden

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