summary
The strong interaction between streptavidin and biotin is one of the most commonly exploited tools in chemistry and biology. Methods for the facile derivatization of a variety of molecules (in particular, proteins) with biotin have been introduced, in order to allow their efficient recovery, immobilization and detection with streptavidin-based reagents. However, when desired, the release of biotinylated proteins from the streptavidin-based reagents remains a major problem, due to the extraordinary stability of this complex. This chapter presents a protocol developed in our laboratory for the quantitative elution of biotinylated proteins from streptavidin sepharose, featuring harsh elution conditions and competition with free biotin. The usefulness of the method is shown by the recovery of biotinylated proteins from organ homogenates, obtained from mice perfused with a reactive ester derivative of biotin.
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Acknowledgments
Financial support of the Gebert-Rüf Foundation, the Swiss National Science Foundation and of the Bundesamt für Bildung und Wissenschaft (EU Projects Angiogenesis and Stroma FPS/6 no. 503233) and the access to instrumentation of the Functional Genomics Center Zurich are gratefully acknowledged. Giuliano Elia is on leave of absence from Institute of Neurobiology and Molecular Medicine CNR, Rome, Italy.
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Rösli, C., Rybak, JN., Neri, D., Elia, G. (2008). Quantitative Recovery of Biotinylated Proteins from Streptavidin-Based Affinity Chromatography Resins. 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_8
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DOI: https://doi.org/10.1007/978-1-59745-579-4_8
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