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Overview of Antibody Use in Immunocytochemistry

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Immunocytochemical Methods and Protocols

Part of the book series: Methods in Molecular Bilogy ((MIMB,volume 34))

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Abstract

Immunocytochemistry, by definition, is the identification of a tissue constituent in situ by means of a specific antigen-antibody interaction where the antibody has been tagged with a visible label (1) Cell staining is a powerful method to demonstrate both the presence and subcellular location of a particular molecule of interest (2) Initial attempts to label antibodies with ordinary dyes were unsatisfactory because the label was not sufficiently visible under the microscope. A. H. Coons first introduced immunofluorescence in 1941, using specific antibodies labeled with a fluorescent dye to localize substances in tissues (3). This technique was considered difficult, and its potential was not widely realized for nearly 20 years. Early attempts focused on labeling the specific antibody itself with a fluorophore (see Chapter 6). The labeled antibody was then applied to the tissue section to identify the antigenic sites (direct method) (3) (see Chapter 15). Later, the more sensitive and versatile indirect method was introduced (4) (see Chapter 16–Chapter 18). In this method, the specific antibody, bound to the antigen, was detected with a secondary reagent, usually another antibody that had been tagged with either a fluorophore or an enzyme.

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References

  1. Van Noorden, S. and Polak, J. M. (1983) Immunocytochemistry today: techniques and practice, in Immunocytochemistry, Practical Applications in Pathology and Biology (Polak, J. M and Van Noorden, S., eds.), Wright PSG, Bristol, England, pp 11–42.

    Google Scholar 

  2. Sternberger, L. A. (1979) Immunocytochemistry, 2nd ed. Wiley, New York.

    Google Scholar 

  3. Coons, A H., Creech, H. J., and Jones, R. N. (1941) Immunological properties of an antibody containing a fluorescent group. Proc. Soc. Exp. iol Med. 47, 200–202

    CAS  Google Scholar 

  4. Coons, A. H, Leduc, E. H, and Connolly, J M. (1955) Studies on antibody production I. A method for the histochemical demonstration of specific antibody and its application to a study of the hyperimmune rabbit. J. Exp. Med. 102, 49–60.

    Article  PubMed  CAS  Google Scholar 

  5. Nakane, P. K. and Pierce, G. Jr. (1966) Enzyme-labeled antibodies: preparation and application for the localization of antigen J. Histochem Cytochem. 14, 929–931

    PubMed  CAS  Google Scholar 

  6. Engvall, E. and Perlman, P. (1971) Enzyme-linked immunosorbent assay (ELISA), Quantitative assay of immunoglobulin G Immunocytochemistry 8, 871–874.

    Article  CAS  Google Scholar 

  7. Massayeff, R and Maillini, R(1975) A sandwich method of enzyme immunoassay. Application to rat and human α-fetoprotein. J. Immunol. Methods 8, 223–234.

    Article  Google Scholar 

  8. Mason, D. Y. and Woolston, R. E. (1982) Double immunoenzymatic labeling, in Techniques in Immunocytochemistry, vol. 1 (Bullock, G. and Petrusz, P, eds), Academic, London, pp. 135–152

    Google Scholar 

  9. Campbell, G T and Bhatnagar, A S. (1976) Simultaneous visualization by light microscopy of two pituitary hormones in a single tissue section using a combination of indirect immunohistochemical methods J Histochem Cytochem 24, 448–452.

    PubMed  CAS  Google Scholar 

  10. Mason, D Y., Abdulaziz, Z, Falini, B and Stein, H (1983) Double immunoenzymatic labeling, in Immunocytochemistry, Practical Applications in Pathology and Biology (Polak, J M and VanNoorden, S., eds.), Wright PSG, Bristol, England, pp. 113–128

    Google Scholar 

  11. Singer, S. J. (1959) Preparation of an electron-dense antibody conjugate. Nature 183, 1523–1524

    Article  PubMed  CAS  Google Scholar 

  12. Faulk, W P and Taylor, G. M (1971) An immunocolloid method for the electron microscope. Immunochemistry 8, 1081–1083.

    Article  PubMed  CAS  Google Scholar 

  13. Roth, J., Bendagan, M., and Orci, L(1978) Ultrastructural localization of intracellular antigens by use of Protein-A gold complex. J. Histochem Cytochem. 26, 1074–1081

    PubMed  CAS  Google Scholar 

  14. Larsson, L.-I and Schwartz, T. W (1977) Radioimmunocytochemistry—a novel immunocytochemical principle. J. Histochem. Cytochem 25, 1140–1146.

    PubMed  CAS  Google Scholar 

  15. Cuello, A C, Priestley, J. V., and Milstein, C. (1982) Immunocytochemistry with internally labeled monoclonal antibodies. Proc Natl. Acad. Sci. USA 78, 665–669.

    Article  Google Scholar 

  16. Livingston, D. M (1974) Immunoaffinity chromatography of proteins. Methods Enzymol. 34, 723–731

    Article  PubMed  CAS  Google Scholar 

  17. Clausen, J. (1981) Immunochemical techniques for the identification and estimation of macromolecules, in Laboratory Techniques in Biochemistry and Molecular Biology, vol. 1, pt 3 (Work, T S. and Work, E, eds.), Elsevier, Amsterdam, pp 52–155.

    Google Scholar 

  18. Brown, R K (1967) Immunological techniques (general). Methods Enzymol 11, 917–927.

    Article  Google Scholar 

  19. Van Regenmortel, M H V., Briand, J P, Muller, S, and Plaué, S. (1988) Immunization with peptides Synthetic peptides as antigens, in Laboratory Techniques in Biochemistry and Molecular Biology, vol 19(Burdon,R H. and vanKnippenberg, P. H., eds.), Elsevier, Amsterdam, pp. 131–158.

    Google Scholar 

  20. Kohler, G. and Milstein, C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256, 495–497.

    Article  PubMed  CAS  Google Scholar 

  21. Galfre G. and Milstein, C(1981) Preparation of monoclonal antibodies: strategies and procedures. Methods Enzymol. 73, 3–46.

    Article  PubMed  CAS  Google Scholar 

  22. Nisonoff, A. (1984) Specificities, affinities, and reaction rates of antihapten antibodies, in Introduction to Molecular Immunology. Sinauer, Sunderland, MA, pp 29–43

    Google Scholar 

  23. Oudin, J (1980) Immunochemical analysis by antigen-antibody precipitation in gels Methods Enzymol. 70, 166–198.

    Article  PubMed  CAS  Google Scholar 

  24. VanVunakis, H (1980) Radioimmunoassays. an overview. Methods Enzymol 70, 201–209.

    Article  PubMed  Google Scholar 

  25. Vandesande, F. (1979) A critical review of immunocytochemical methods for light microscopy. J. Neurosci. Methods 1, 3–23.

    Article  PubMed  CAS  Google Scholar 

  26. Grube, D. (1980) Immunoreactivities of gastrin (G) cells II Nonspecific binding of immunoglobulins to G-cells by ionic interactions. Histochemistry 66, 149–167.

    Article  PubMed  CAS  Google Scholar 

  27. DeMey, J. and Moeremans, M. (1986) Raising and testing polyclonal antibodies for immunocytochemistry, in Immunocytochemistry: Modern Methods and Applications (Polak, J M and VanNoorden, S., eds), Wright, Bristol, England, pp. 3–12.

    Google Scholar 

  28. Ritter, M A. (1986) Raising and testing monoclonal antibodies for immunocytochemistry, in Immunocytochemistry Modern Methods and Applications (Polak, J. M. and VanNoorden, S, eds.), Wright, Bristol, England, pp. 13–25.

    Google Scholar 

  29. VanNoorden, S. (1986) Tissue preparation and immunostaining techniques for light microscopy, in Immunocytochemistry: Modern Methods and Applications (Polak, J. M. and VanNoorden, S., eds.), Wright, Bristol, England, pp. 26–53.

    Google Scholar 

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Authors and Affiliations

  1. National Institute of Arthritis & Musculoskeletal & Skin Diseases, National Institutes of Health, Bethesda, MD

    Su- Yau Mao & Ute M. Kent

  2. Department of Biology, The Catholic University of America, Washington, DC

    Lorette C. Javois

Authors
  1. Su- Yau Mao
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  2. Lorette C. Javois
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  3. Ute M. Kent
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Editor information

Editors and Affiliations

  1. The Catholic University of America, Washington, DC

    Lorette C. Javois

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© 1994 Humana Press Inc.

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Yau Mao, S., Javois, L.C., Kent, U.M. (1994). Overview of Antibody Use in Immunocytochemistry. In: Javois, L.C. (eds) Immunocytochemical Methods and Protocols. Methods in Molecular Bilogy, vol 34. Humana Press. https://doi.org/10.1385/0-89603285-X:3

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  • DOI: https://doi.org/10.1385/0-89603285-X:3

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-285-9

  • Online ISBN: 978-1-59259-521-1

  • eBook Packages: Springer Protocols

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