Abstract
Biotin—avidin detection systems are widely used in both immunocytochemistry and molecular biology (1,2) (see Chapter 25). They take advantage of the high affinity of biotin, a low-molecular-weight vitamin, for avidin, an egg-white protein. The avidin-biotin complex has one of the highest dissociation constants known, 10-15 M. This high dissociation constant has made it a convenient system for linking indicators to antibodies. However, egg-white avidin binds nonspecifically to many tissue sites. This nonspecific binding has been attributed both to its high isoelectric point (pI =10) and to the fact that the protein is glycosylated (3,4). Because of the problem of nonspecific binding of avidin, streptavidin has largely replaced avidin for immunocytochemical procedures. Streptavidin, produced by Streptomyces avidinni, has properties that are very similar to avidin, but it is not glycosylated (5). Streptavidin generally gives little or no background staining, and is superior to avidin for immunocytochemical uses.
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References
Bonnard, C., Papermaster, D. S., and Kraehenbuhl, J.-P. (1984) The streptavidinbiotin bridge technique: application in light and electron microscope immunocytochemistry, in Immunolabelling for Electron Microscopy (Polak, J. M. and Varndell, I. M., eds.), Elsevier, New York, pp. 95–111.
Larsson, L.-I. (1988) Immunocytochemical detection systems, in Immunocytochemistry: Theory and Practice (Larsson, L.-I., ed.), CRC, Boca Raton, FL, pp.77–146.
Wooley, D. W. and Longsworth, L. G. (1942) Isolation of an antibiotin factor from egg white. J. Biol. Chem. 142, 285–290.
Green, N. M. (1975) Avidin. Adv. Prot. Res. 29, 85–133.
Chaiet, L. and Wolf, F. J. (1964) The properties of streptavidin, a biotin-binding protein produced by Streptomycetes. Arch. Biochem. Biophys. 106, 1–5.
Reynolds, E. S. (1963) The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J. Cell Biol. 17, 208–212.
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© 1999 Humana Press Inc., Totowa, NJ
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Oliver, C. (1999). Colloidal Gold/Streptavidin Methods. In: Javois, L.C. (eds) Immunocytochemical Methods and Protocols. Methods in Molecular Biology™, vol 115. Humana Press. https://doi.org/10.1385/1-59259-213-9:335
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DOI: https://doi.org/10.1385/1-59259-213-9:335
Publisher Name: Humana Press
Print ISBN: 978-0-89603-813-4
Online ISBN: 978-1-59259-213-5
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