Advertisement

The Histochemical Journal

, Volume 20, Issue 2, pp 75–80 | Cite as

Improved avidin—biotin—peroxidase complex (ABC) staining

  • Giorgio Cattoretti
  • Emilio Berti
  • Raffaella Schiró
  • Lucia D'Amato
  • Costante Valeggio
  • Franco Rilke
Papers

Summary

A considerable intensification of the avidin—biotin—peroxidase complex staining system (ABC) was obtained by sequentially overlaying the sections to be immunostained with an avidin-rich and a biotin-rich complex. Each sequential addition contributed to the deposition of horseradish peroxidase on the immunostained site and allowed the subsequent binding of a complementary complex. With this technique a higher dilution of the antisera could be used and minute amounts of antigen masked by the fixative could be demonstrated on paraffin sections.

Keywords

Paraffin Biotin Horseradish Peroxidase Paraffin Section Sequential Addition 
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.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. CORDELL, J. L., FALINI, B., ERBER, W. N., GHOSH, A. K., ABDULAZIZ, Z., MACDONALD, S., PULFORD, K. A. F., STEIN, H. & MASON, D. Y. (1984) Immunoenzymatic labelling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes).J. Histochem. Cytochem. 52, 219–29.Google Scholar
  2. DEPETRIS, S. (1982) Immunoelectron microscopy and immunofluorescence in membrane biology. InMethods in Membrane Biology, Vol. 9 (edited by KORN, E. D.) pp. 1–200. New York: Plenum.Google Scholar
  3. GUPTA, P. K., MYERS, J. D., BAYLIN, S. B., MULSHINE, J. L., CUTITTA, F. & GAZDAR, A. F. (1985) Improved antigen detection in ethanol-fixed cytologic specimens. A modified Avidin-Biotin-Peroxidase complex (ABC) method.Diagn. Cytopathol. 1, 133–6.Google Scholar
  4. HSU, S., RAINE, L. & FANGER, H. (1981a) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabelled antibody (PAP) procedures.J. Histochem. Cytochem. 29, 577–80.Google Scholar
  5. HSU, S., RAINE, L. & FANGER, H. (1981b) The use of anti-avidin antibody and avidin-biotin-peroxidase complex in immunoperoxidase technics.Amer. J. Clin. Pathol. 75, 816–21.Google Scholar
  6. LANSDORP, P. M., VAN DER KWAST, T. H., DE BOER, M. & ZEIJLEMAKER, W. P. (1981) Stepwise amplified immunoperoxidase (PAP) staining. Cellular morphology in relation to membrane markers.J. Histochem. Cytochem. 52, 172–8.Google Scholar
  7. MADRI, J. A. & BARWICH, K. W. (1983) Use of avidinbiotin complex in an ELISA system: a quantitative comparison with two other immunoperoxidase detection systems using keratin antisera.Lab. Invest. 48, 98–107.Google Scholar
  8. VACCA, L. L. (1982) ‘Double bridge’ techniques of immunocytochemistry. InTechniques in Immunocytochemistry, Vol. 1 (edited by BULLOCK, G. R. and PETRUSZ, P.) pp. 155–82. London: Academic Press.Google Scholar
  9. WOOD, G. S. & WARNKE, R. (1981) Suppression of endogenous avidin-binding activity in tissues and its relevance to biotin-avidin detection system.J. Histochem. Cytochem. 29, 1196–2004.Google Scholar

Copyright information

© Chapman and Hall Ltd 1988

Authors and Affiliations

  • Giorgio Cattoretti
    • 1
  • Emilio Berti
    • 2
  • Raffaella Schiró
    • 3
  • Lucia D'Amato
    • 1
  • Costante Valeggio
    • 3
  • Franco Rilke
    • 1
  1. 1.Division of Anatomic Pathology and CytologyIstituto Nazionale per lo Studio e la Cura dei TumoriMilanoItaly
  2. 2.1st Department of DermatologyUniversitá Statale di MilanoMilanoItaly
  3. 3.Laboratorio di Ricerche ClinicheIstituti Clinici di PerfezionamentoMilanoItaly

Personalised recommendations