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Purification of biotinylated proteins on streptavidin resin: A protocol for quantitative elution
Author(s) -
Rybak JaschaN.,
Scheurer Simone B.,
Neri Dario,
Elia Giuliano
Publication year - 2004
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200300780
Subject(s) - biotinylation , streptavidin , biotin , elution , chromatography , chemistry , sepharose , biochemistry , enzyme
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.