Skip to main content

Enzyme and Biotin Labeling of Antibody

  • Protocol
Basic Protein and Peptide Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 32))

Abstract

Labeled antibodies are key reagents in many procedures, particularly immunoassays and the detection of antigens in tissue sections and on membrane blots. The ideal labeled antibody retains all the immunoreactivity of the antibody and all the signal-generating capacity of the label. In addition, these conjugates should be stable, and have defined and appropriate proportions of the two components.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Tijssen, P. (1985) Practice and Theory of Enzyme Immunoassays, Elsevier, Amsterdam, pp. 221–278.

    Google Scholar 

  2. Ishikawa, E., Hashida, S., Kohno, T., and Tanaka, K. (1988) Methods for enzyme labeling of antigens, antibodies and their fragments, in Nonisotopic Immunoassay (Ngo, T. T., ed.), Plenum, New York, pp. 27–55.

    Google Scholar 

  3. Engvall, E. and Perlmann, P. (1972) Enzyme-linked immunosorbent assay, ELISA. III. Quantitation of specific antibodies by enzyme-labelled anti-immunoglobulin in antigen-coated tubes. J. Immunol. 109, 129–135.

    PubMed  CAS  Google Scholar 

  4. Wilson, M. B. and Nakane, P. P. (1978) Recent developments in the periodate method of conjugating horseradish peroxidase (HRPO) to antibodies, in Immunofluorescence and Related Staining Techniques (Knapp, W., Holubar, K., and Wick, G., eds.), Elsevier/North Holland Biomedical, Amsterdam, pp. 215–224.

    Google Scholar 

  5. O’Sullivan, M. J., Gnemmi, E., Morris, D., Chieregatti, G., Simmonds, A. D., Simmons, M., Bridges, J. W., and Marks, V. (1979) Comparison of two methods of preparing enzyme-antibody conjugates: Application of these conjugates to enzyme immunoassay. Anal. Biochem. 100, 100–108.

    Article  PubMed  Google Scholar 

  6. Carlsson, J., Drevin, H., and Axen, R. (1978) Protein thiolation and reversible protein-protein conjugation, N-succinimidyl 3-(2-pyridyldithio)propionate—a new heterobifunctional reagent. Biochem. J. 173, 723–737.

    PubMed  CAS  Google Scholar 

  7. Halliday, M. I. and Wisdom, G. B. (1986) A comparison of three methods for the preparation of enzyme antibody conjugates. Biochem. Soc. Trans. 14, 473–474.

    CAS  Google Scholar 

  8. Diamandis, E. P. and Christpoulos, T. K. (1991) The biotin-(strept)avidin system: principles and applications in biotechnology. Clin. Chem. 37, 625–636.

    PubMed  CAS  Google Scholar 

  9. Guesdon, J.-L., Ternynck, T., and Avrameas, S. (1979) The use of avidin-biotin interaction in immunoenzymatic techniques. J. Histochem. Cytochem. 27, 1131–1139.

    PubMed  CAS  Google Scholar 

  10. Bayer, E. A. and Wilchek, M. (1980) The use of avidin-biotin complex as a tool in molecular biology. Methods Biochem. Anal. 26, 1–45.

    Article  PubMed  CAS  Google Scholar 

  11. Ellman, G. L. (1959) Tissue sulfhydryl groups. Arch. Biochem. Biophys. 82, 70–75.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1994 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Wisdom, G.B. (1994). Enzyme and Biotin Labeling of Antibody. In: Walker, J.M. (eds) Basic Protein and Peptide Protocols. Methods in Molecular Biology™, vol 32. Humana Press. https://doi.org/10.1385/0-89603-268-X:433

Download citation

  • DOI: https://doi.org/10.1385/0-89603-268-X:433

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-268-2

  • Online ISBN: 978-1-59259-519-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics