Journal
PROTEOMICS
Volume 4, Issue 8, Pages 2296-2299Publisher
WILEY
DOI: 10.1002/pmic.200300780
Keywords
affinity chromatography; avidin; biotin; biotinylation; streptavidin
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The interaction between streptavidin and biotin is one of the most widely used tools in chemistry and biology. However, the release of biotinylated proteins from streptavidin resins remains a major problem, due to the extraordinary stability of this complex. We present a new protocol for the quantitative elution of biotinylated proteins from streptavidin Sepharose, featuring harsh elution conditions and competition with free biotin. The usefulness of the method was demonstrated by the quantitative recovery of biotinylated proteins from organ homogenates, obtained from mice perfused with a reactive ester derivative of biotin.
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