• Kari Johansen
  • Lennart Svensson
Part of the Methods in Molecular Medicine™ book series (MIMM, volume 13)


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, 2, 3).


Ammonium Persulfate Bromphenol Blue Radiolabeled Amino Acid Detergent Lysis Immunoprecipitation Technique 
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, NYGoogle Scholar
  2. 2.
    Harlow E and Lane D (1988) Antibodies. A Laboratory Manual Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NYGoogle Scholar
  3. 3.
    Burleson F G, Chambers T. M, and Wiedbrauk D L (eds) (1992) Virology A Laboratory Manual Academic, San Diego, CAGoogle Scholar
  4. 4.
    Barrett A. J and Salvesen G (eds.) (1986) Proteinase Inhibitors Research Monographs in Cell and Tissue Physiology, vol. 12. Elsevler, Amsterdam, The NetherlandsGoogle Scholar
  5. 5.
    James G. T (1978) Inactivation of the protease inhibitor phenylmethylsulfonyl fluoride in buffers Anal. Biochem 86, 574–579PubMedCrossRefGoogle 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, 41–52PubMedCrossRefGoogle Scholar
  7. 7.
    Hjelm H, Hjelm K, and Sjoquist J. (1972) Protein A from Staphyiococcus aureus Its isolation by affinity chromatography and its use as an immunosorbent for isolation of immunoglobulins FEBS Lett 28, 73–76PubMedCrossRefGoogle Scholar
  8. 8.
    Sjodahl J. (1977) Structural studies on the four repetitive Fc-binding regions in protein A from Staphylococcus aureus Eur J Biochem 78, 471–490PubMedCrossRefGoogle 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, 21–28PubMedCrossRefGoogle Scholar
  10. 10.
    Bjorck L. and Kronvall G (1984) Purification and some properties of strepto-coccal protein G a novel IgG-binding reagent J Immunol 133, 969–974PubMedGoogle Scholar
  11. 11.
    Åkerstrom B, Brodin T, Reis K., and Bjórck L (1985) Protein G: a powerful tool for binding and detection of monoclonal and polyclonal antibodies J Immunol 135, 2589–2592.PubMedGoogle Scholar
  12. 12.
    Fahnestock S. R., Alexander P, Nagle J., Filpula D (1986) Gene for an immunoglobulin-binding protein from a group G streptococcus J Bacterial 167(3), 870–880Google Scholar
  13. 13.
    Roque-Barreira M C and Campos-Neto A (1985) Jacalin: an IgA-binding lectin J Immunol. 134, 1740–1743PubMedGoogle Scholar
  14. 14.
    Johansen K., Granqvist L., Karlén 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 Mel Biol 79, 237–248CrossRefGoogle Scholar
  16. 16.
    Hames B. D. and Rickwood D. (eds.) (1981) Gel Electrophoresis of Proteins A Practical Approach IRL, Oxford, UKGoogle Scholar
  17. 17.
    Omstein L (1964) Disc electrophoresis-I Background and theory Ann NY Acad Sci 121, 321–349Google Scholar
  18. 18.
    Davis B J (1964) Disc-electrophoresis II. Method and application to human serum proteins. Ann NY Acad Sci 121, 404–427PubMedCrossRefGoogle Scholar
  19. 19.
    Laemmli U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 1998

Authors and Affiliations

  • Kari Johansen
    • 1
  • Lennart Svensson
    • 1
  1. 1.Department of VirologySwedish Institute for Infectious Diseases ControlStockholmSweden

Personalised recommendations