Biotinylation of Antibodies

  • Su-Yau Mao
Part of the Methods in Molecular Biology™ book series (MIMB, volume 115)


The high affinity and specificity of the avidin-biotin interaction permit diverse applications in immunology, histochemistry, in situ hybridizations, affinity chromatography, and many other areas (1) (see Chapters 25 and 26). It was first exploited in immunocytochemical applications in the mid-1970s (2,3), and has since been commonly used to localize antigens in cells and tissues. In this technique, a biotinylated primary or secondary antibody is first applied to the sample, and the detection is accomplished by using labeled avidin. Avidin with a variety of labels are available commercially, including fluorescent, enzyme, iodine, ferritin, or gold labels.


Sodium Azide Gold Label Sodium Borate Buffer Charge Cell Surface Increase Protein Concentration 
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.
    Roffman, E., Meromsky, L., Ben-Hur, H., Bayer, E. A., and Wilchek, M. (1986) Selective labeling of functional groups on membrane proteins or glycoproteins using reactive biotin derivatives and 125I-streptavidin. Biochem. Biophys. Res. Comm. 136, 80–85.PubMedCrossRefGoogle Scholar
  2. 2.
    Becker, J. M. and Wilchek, M. (1972) Inactivation by avidin of biotin-modified bacteriophage. Biochim. Biophys. Acta 264, 165–170.PubMedGoogle Scholar
  3. 3.
    Heitzmann, H. and Richards, F. M. (1974) Use of the biotin-avidin complex for specific staining of biological membranes in electron microscopy. Proc. Natl. Acad. Sci. USA 71, 3537–3541.PubMedCrossRefGoogle Scholar
  4. 4.
    Green, N. M. (1975) Avidin. Adv. Protein Chem. 29, 85–133.PubMedCrossRefGoogle Scholar
  5. 5.
    Hsu, S.-M., Raine, L., and Fanger, H. (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques. J. Histochem. Cytochem. 29, 577–580.PubMedGoogle Scholar
  6. 6.
    Guesdon, J. L., Ternynck, T., and Avrameas, S. (1979) The use of avidin-biotin inter action in immuno-enzymatic techniques. J. Histochem. Cytochem. 27, 1131–1139.PubMedGoogle Scholar
  7. 7.
    Lee, W. T. and Conrad, D. H. (1984) The murine lymphocyte receptor for IgE. II. Characterization of the multivalent nature of the B lymphocyte receptor for IgE. J. Exp. Med. 159, 1790–1795.PubMedCrossRefGoogle Scholar
  8. 8.
    LaRochelle, W. J. and Froehner, S. C. (1986) Determination of the tissue distributions and relative concentrations of the postsynaptic 43-kDa protein and the acetylcholine receptor in Torpedo. J. Biol. Chem. 261, 5270–5274.Google Scholar
  9. 9.
    Suter, M. and Butler, J. E. (1986) The immunochemistry of sandwich ELISAs. II. A novel system prevents the denaturation of capture antibodies. Immunol. Lett. 13, 313–316.PubMedCrossRefGoogle Scholar
  10. 10.
    O′Shannessy, D. J., Dobersen, M. J., and Quarles, R. H. (1984) A novel procedure for labeling immunoglobulins by conjugation to oligosaccharide moieties. Immunol. Lett. 8, 273–277.PubMedCrossRefGoogle Scholar
  11. 11.
    O′Shannessy, D. J. and Quarles, R. H. (1987) Labeling of the oligosaccharide moieties of immunoglobulins. J. Immunol. Methods 99, 153–161.PubMedCrossRefGoogle Scholar
  12. 12.
    Bayer, E. A., Zalis, M. G., and Wilchek, M. (1985) 3-(N-Maleimido-propionyl)-biocytin: a versatile thiol-specific biotinylating reagent. Anal. Biochem. 149, 529–536.PubMedCrossRefGoogle Scholar
  13. 13.
    Sutoh, K., Yamamoto, K., and Wakabayashi, T. (1984) Electron microscopic visualization of the SH1 thiol of myosin by the use of an avidin-biotin system. J. Mol. Biol. 178, 323–339.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1999

Authors and Affiliations

  • Su-Yau Mao
    • 1
  1. 1.Medimmune, Inc.Gaithersburg

Personalised recommendations