Immunoperoxidase Techniques Using the Avidin-Biotin System

  • Su-Ming Hsu


Although immunofluorescence techniques were first develpoed in the early 1960s, they did not gain widespread use and acceptance until the following decade, when they became invaluable tools for the resolution of a number of problems in experimental immunology. Since then, the method has been used clinically for the identification of surface and intracellular lymphocyte markers and immune complexes of glomerular diseases. Within the past 15 years, an alternative method using enzyme-labeled antibodies has been developed which is rapidly replacing immunofluorescence because of its simplicity and usefulness in formalin-fixed paraffin embedded tissue sections. Although many enzymes have been sucessfully conjugated to various antibodies, most recent studies have employed horseradish peroxidase. The amount of antigens in tissue sections is generally determined semi-quantitatively by examining the staining intensity using serial diluted antibodies (Hsu et al., 1981a,b). More recently, the immunoperoxidase techniques have been adapted to other solid phase systems, such as enzyme-linked immunoassys or western blots of antigens (Azen and Yu, 1984; Madri and Barwick, 1983; Rosenkoetter et al., 1984).


Clin Pathol Immunoperoxidase Technique Peanut Agglutinin Solid Phase System Avidin Molecule 
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  1. Azen, E.A., Yu, P.L., 1984, Genetics polymorphism of CON 1 and CON 2 salivary proteins detected by immunologic and Con-canavalin A reactions on nitrocellulose with linkage of CON 1 and CON 2 genes to the SPC (salivary protein gene complex), Biochem Genetics 22:19.Google Scholar
  2. Bayer, E.A., Wilcheck, M., 1980, The use of the avidin-biotin complex as a tool in molecular biology, Methods Biochem Anal 26:1.PubMedCrossRefGoogle Scholar
  3. Childs, G., Unabia, G., 1982, The application of the avidin-biotin-peroxidase complex (ABC) method to the light microscopic localization of pituitary hormones, J Histochem Cytochem 30:713.PubMedCrossRefGoogle Scholar
  4. Childs, G., Unabia, G., 1982, Application of a rapid avidin-biotin peroxidase complex (ABC) technique to the localization of pituitary hormones at the electron microscopic level, J Histochem Cytochem, 30:1320PubMedCrossRefGoogle Scholar
  5. Guesdon, J.L., Ternynck, T., Avrameas, S., 1979, The use of avidin-biotin interaction in immunoenzymatic techniques. J Histochem Cytochem 27:1131.PubMedCrossRefGoogle Scholar
  6. Hofmann, K., Kiso, Y., 1976, An approach to the targeted attachment of peptides and proteins to solid supports. Proc Natl Acad Sci USA 73:3516.PubMedCrossRefGoogle Scholar
  7. Hofmann, K., Finn, F.M., Friesen, H.J., Diaconescu, C., Zahn, H.,1977, Biotinyl insulins as potential tools for receptor studies. Proc Natl Acad Sci USA 74:2697PubMedCrossRefGoogle Scholar
  8. Holland, P.C., Pena, S.D.J., Guerin, C.W., 1984, Developmental regulation of neuramidase-sensitive lectin binding glycoproteins during myogenesis of rat L6 myoblasts. Biochem J 218:465.PubMedGoogle Scholar
  9. Hsu, P.L., Hsu, S.M., and Appela, E., 1985, Immunological screening of a cDNA library using avidin-biotin-peroxidase complex method. Gene Anal Tech, in press.Google Scholar
  10. Hsu, S.M., Raine, L., Fänger, H., 1981a, A comparative study of the peroxidase-anti-peroxidase method and an avidin-biotin complex method for studying polypeptide hormones with radioimmunoassay antibodies. Am J Clin Pathol 75:734.PubMedGoogle Scholar
  11. Hsu, S.M., Raine, L., Fanger, H., 1981b, Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: A comparison between ABC & unlabeled antibody (PAP) procedures. J Histochem Cytochem 29:577.PubMedCrossRefGoogle Scholar
  12. Hsu, S.M., Raine, L., 1981c, Protein A, avidin and biotin in immunohistochemistry. J Histochem Cytochem 29:1349.PubMedCrossRefGoogle Scholar
  13. Hsu, S.M., Raine, L., 1982a, Discovery of high affinity of Pha-seolus vulgaris agglutinin (PHA) with gastrin-secreting cells. Am J Clin Pathol 77:396.PubMedGoogle Scholar
  14. Hsu, S.M., Raine, L., 1982b, Unwanted binding associated with avidin-biotin-peroxidase complex in frozen tissues. Am J Clin Pathol 77:775.Google Scholar
  15. Hsu, S.M., Raine, L., 1982c, Versatility of biotin-labeled lectins and avidin-biotin-peroxidase complex for localization of carbohydrates in tissue sections. J Histochem Cytochem 30:157.PubMedCrossRefGoogle Scholar
  16. Hsu, S.M., Raine, L., 1982d, Warthin’s tumor-epithelial cell differences. Am J Clin Pathol 77:78.PubMedGoogle Scholar
  17. Hsu, S.M., Raine, L., 1982e, Concanavalin A (Con A) reaction with plasma membrane. Am J Clin Pathol 77:508.PubMedGoogle Scholar
  18. Hsu, S.M., Soban, E., 1982, Color modification of diaminobenzi-dine (DAB) precipitation by metallic ions and its applications for double immunohistochemistry. J Histochem Cytochem 30:1079.PubMedCrossRefGoogle Scholar
  19. Hsu, S.M., Ree, H., 1983, Histochemical studies on lectin binding in reactive lymphoid tissues. J Histochem Cytochem 31:538PubMedCrossRefGoogle Scholar
  20. Hsu, S.M., Raine, L., 1983, The use of avidin-biotin-peroxidase-complex (ABC) in diagnostic and research pathology. In Advances in Immunohistochemistry. Ed: R.A. DeLellis, Masson Publishing USA, Inc., New York, pp 31–42.Google Scholar
  21. Leary, J.J., Brigati, D.J., and Ward, D.C., 1983, Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bio-blots. Proc Natl Acad Sci USA 80:4045.PubMedCrossRefGoogle Scholar
  22. Leipold, B., Remy, W., 1984, Use of avidin-biotin peroxidase complex for measurement of UV lesions in human DNA by micro-ELISA. J Immuno Methods 66:227.CrossRefGoogle Scholar
  23. Madri, J.A., Barwick, K.W., 1983, Use of avidin-biotin-complex in ELISA system. A quantitative comparison with two.other immunoperoxidase detection systems using keratin antisera. Lab Invest 48:98PubMedGoogle Scholar
  24. Myerson, D., Hackman, R.C., Meyers, J.D., 1984, Diagnosis of cytomegaloviral pneumonia by in situ hybridization. J Infect Diseases. 150:272.CrossRefGoogle Scholar
  25. Rosenkoetter, M., Reder, A.T., Oger, J.J.F., Antel, J.P., 1984, T cell regualtion of polyclonally induced immunoglobulin secretion in humans. J Immunol 132:1779.PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1985

Authors and Affiliations

  • Su-Ming Hsu
    • 1
  1. 1.Department of Pathology and Laboratory MedicineUniversity of Texas Health Science CenterHoustonUSA

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