Advertisement

Conjugation of Colloidal Gold to Proteins

  • Constance Oliver
Part of the Methods in Molecular Biology book series (MIMB, volume 588)

Abstract

The ability to conjugate proteins to colloidal gold sols provides a wide variety of probes for electron microscopy. Antibodies, protein A, protein G, lectins, enzymes, toxins, and other proteins have all been conjugated to colloidal gold. The nature of the interaction between the colloidal gold and the protein is poorly understood. Proteins are conjugated to gold sols by adjusting the pH of the gold sol to approximately 0.5 pH unit higher than the pI of the protein being conjugated. This chapter gives a general method for conjugating proteins to colloidal gold as well as more specific methods for conjugating antibodies and protein A to colloidal gold.

Key words

Antibody Protein A Protein G Conjugation Colloidal gold 

References

  1. 1.
    Leunissen JLM, DeMey JR (1989) Preparation of gold probes. In: Verkleij AJ, Leunissen JLM (eds) Immuno-gold Labeling in Cell Biology. CRC, Boca Raton, FL, pp 3–16Google Scholar
  2. 2.
    Bendayan M (1984) Protein A-gold electron microscopic immunocytochemistry: methods, applications, and limitations. J Electron Micros Technol 1:243–270CrossRefGoogle Scholar
  3. 3.
    Bendayan M (1989) Protein A-gold and protein G-gold postembedding immunoelectron microscopy. In: Hayat MA (ed) Colloidal Gold, vol. 1. Academic Press, New York, NY, pp 34–96Google Scholar
  4. 4.
    Horisberger M (1981) Colloidal gold: a cytochemical marker for light and fluorescent microscopy and for transmission and scanning electron microscopy. In: Johari O (ed) Scanning Electron Microscopy II. SEM, Inc., AMF O’Hare, Chicago, IL, pp 9–31Google Scholar
  5. 5.
    Horisberger M (1989) Quantitative aspects of labeling colloidal gold with proteins. In: Verkleij AJ, Leunissen JLM (eds) Immuno-gold Labeling in Cell Biology. CRC, Boca Raton, FL, pp 49–60Google Scholar
  6. 6.
    Geoghegan WD, Ackerman GA (1977) Adsorption of horseradish peroxidase, ovomucoid and anti-immunoglobulin to colloidal gold for the indirect detection of concanavalin A, wheat germ agglutinin and goat anti-human immunoglobulin G on cell surfaces at the electron microscopic level: a new method, theory and application. J Histochem Cytochem 25:1187–1200PubMedGoogle Scholar
  7. 7.
    Roth J (1983) The colloidal gold marker system for light and electron microscopic cytochemistry. In: Bullock GR, Petrusz P (eds) Techniques in Immunocytochemistry, vol. 2. Academic Press, New York, NY, pp 217–284Google Scholar
  8. 8.
    Birrell GB, Hedberg KK, Griffith PH (1987) Pitfalls of immunogold labeling: analysis by light microscopy, transmission electron microscopy, and photoelectron microscopy. J Histochem Cytochem 35:843–853PubMedGoogle Scholar
  9. 9.
    Gasparyan VK (2005) Hen egg immunoglobulin Y in colloidal gold agglutination assay: Comparison with rabbit immunoglobulin G. J Clin Lab Anal 19:124–127CrossRefPubMedGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Department of Cell and Molecular Biology and Pathogenic Bioagents, Faculdade de Medicina de Ribeirão PretoUniversity of São PauloRibeirão PretoBrazil

Personalised recommendations