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Electrophoretic behavior of streptavidin complexed to a biotinylated probe: A functional screening assay for biotin-binding proteins

Humbert, Nicolas and Zocchi, Andrea and Ward, Thomas R.. (2005) Electrophoretic behavior of streptavidin complexed to a biotinylated probe: A functional screening assay for biotin-binding proteins. Electrophoresis, 26 (1). pp. 47-52.

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Official URL: http://edoc.unibas.ch/dok/A5254451

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Abstract

The biotin-binding protein streptavidin exhibits a high stability against thermal denaturation, especially when complexed to biotin. Herein we show that, in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), streptavidin is stabilized at high temperature in the presence of biotinylated fluorescent probes, such as biotin-4-fluorescein, which is incorporated within the binding pocket. In nondenaturing SDS-PAGE, streptavidin is detectable when complexed with biotin-4-fluorescein using a UV-transilluminator. Using biotin-4-fluorescein, the detection limit of streptavidin lies in the same range as with Coomassie blue staining. The functionality of streptavidin mutants can readily be assessed from crude bacterial extracts using biotin-4-fluorescein as a probe in nondenaturing SDS-PAGE.
Faculties and Departments:05 Faculty of Science > Departement Chemie > Chemie > Bioanorganische Chemie (Ward)
UniBasel Contributors:Ward, Thomas R. R.
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:Wiley
ISSN:0173-0835
e-ISSN:1522-2683
Note:Publication type according to Uni Basel Research Database: Journal article
Identification Number:
Last Modified:24 Apr 2017 13:16
Deposited On:22 Mar 2012 14:06

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