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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
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 3–16
Bendayan M (1984) Protein A-gold electron microscopic immunocytochemistry: methods, applications, and limitations. J Electron Micros Technol 1:243–270
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 34–96
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 9–31
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 49–60
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 25:1187–1200
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 217–284
Birrell GB, Hedberg KK, Griffith PH (1987) Pitfalls of immunogold labeling: analysis by light microscopy, transmission electron microscopy, and photoelectron microscopy. J Histochem Cytochem 35:843–853
Gasparyan VK (2005) Hen egg immunoglobulin Y in colloidal gold agglutination assay: Comparison with rabbit immunoglobulin G. J Clin Lab Anal 19:124–127
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Oliver, C. (2010). Conjugation of Colloidal Gold to Proteins. In: Oliver, C., Jamur, M. (eds) Immunocytochemical Methods and Protocols. Methods in Molecular Biology, vol 588. Humana Press. https://doi.org/10.1007/978-1-59745-324-0_39
Download citation
DOI: https://doi.org/10.1007/978-1-59745-324-0_39
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-58829-463-0
Online ISBN: 978-1-59745-324-0
eBook Packages: Springer Protocols