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Detection of Biotinylated Proteins in Polyacrylamide Gels Using an Avidin–Fluorescein Conjugate

✍ Scribed by Michihiro Nakamura; Kouhei Tsumoto; Kazunori Ishimura; Izumi Kumagai

Publisher
Elsevier Science
Year
2002
Tongue
English
Weight
97 KB
Volume
304
Category
Article
ISSN
0003-2697

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✦ Synopsis

Biotinylated proteins are widely used as a molecular tool in biotechnological applications. In this paper, we demonstrated that biotinylated proteins after electrophoresis were detected directly in gels using an avidin-fluorescein conjugate with a fluorescence image analyzer. Upon analysis of the purified and chemically biotinylated protein, the sensitivity of this method was almost equal to that of silver staining. Chemically biotinylated proteins of Escherichia coli cell surfaces could also be specifically detected with our method. Furthermore, recombinant proteins fused with the biotin acceptor domain and biotinylated enzymatically in vivo were also detected in a lysate of E. coli specifically. The sensitivity and specificity of our method are high, and the procedure is simple. Therefore, our method would benefit detection of biotinylated proteins via gel electrophoresis and also various fields of study using avidin-biotin technology.

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