summary
Numerous methods have been published for the detection of protein using avidin–biotin technology. Complications can arise using this system when the protein of interest is in extremely high or low abundance. The ability to successfully detect high- or low-abundance proteins is dependent on the detection system selected. The expression of endogenous biotinylated proteins in rat liver homogenate will be used to illustrate the methods utilized for the detection of high- and low-abundance proteins on a membrane. The advantages and disadvantages of enzymatic and direct fluorescence detection systems are discussed.
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References
Neumaier, M., Fenger, U., and Wagener, C. (1986) Anal Biochem 156, 76–80.
Dunn, M. J. (1994) Methods Mol Biol 32, 227–32.
Ruggiero, F. P., and Sheffield, J. B. (1998) J Histochem Cytochem 46, 177–83.
Magnusson, S., Hou, M., Hallberg, E. C., Breimer, M. E., and Wadenvik, H. (1998) Thromb Res 89, 53–8.
Goldman, E. R., Balighian, E. D., Mattoussi, H., Kuno, M. K., Mauro, J. M., Tran, P. T., and Anderson, G. P. (2002) J Am Chem Soc 124, 6378–82.
Khosravi, M. J., and Morton, R. C. (1991) Clin Chem 37, 58–63.
Hitt, A. L., Laing, S. D., and Olson, S. (2002) Anal Biochem 310, 67–71.
Martin, K., Hart, C., Schulenberg, B., Jones, L., and Patton, W. F. (2002) Proteomics 2, 499–512.
Lewis, B., Rathman, S., and McMahon, R. J. (2003) J Nutr Biochem 14, 196–202.
Praul, C. A., Brubaker, K. D., Leach, R. M., and Gay, C. V. (1998) Biochem Biophys Res Commun 247, 312–4.
Lewis, B., Rathman, S., and McMahon, R. (2001) J Nutr 131, 2310–5.
Acknowledgments
The authors thank Dr. Robert J. McMahon for his support, advice, and encouragement of the project.
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© 2008 Humana Press, a part of Springer Science+Business Media, LLC
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Zwart, S.R., Lewis, B.J. (2008). Optimization of Detection and Quantification of Proteins on Membranes in Very High and Very Low Abundance Using Avidin and Streptavidin. In: McMahon, R.J. (eds) Avidin-Biotin Interactions. Methods In Molecular Biology™, vol 418. Humana Press. https://doi.org/10.1007/978-1-59745-579-4_3
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DOI: https://doi.org/10.1007/978-1-59745-579-4_3
Publisher Name: Humana Press
Print ISBN: 978-1-58829-583-5
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