Ultrabright fluorescein-labeled antibodies near silver metallic surfaces |
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Authors: | Lakowicz Joseph R Malicka Joanna Huang Jun Gryczynski Zygmunt Gryczynski Ignacy |
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Institution: | Center for Fluorescence Spectroscopy, University of Maryland at Baltimore, Department of Biochemistry, 725 West Lombard Street, Baltimore, MD 21201, USA. lakowicz@cfs.umbi.umd.edu |
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Abstract: | Fluorescein-labeled antibodies are widely used in clinical assays and fluorescence microscopy. The fluorescent signal per labeled antibody is limited by fluorescein self-quenching, which occurs when the antibody is heavily labeled with multiple fluoresceins. We examined immunoglobulin G (IgG) when labeled with 0.7 to about 30 fluoresceins per antibody molecule. The extent of self-quenching was decreased, and the signal increased, when the labeled antibody was in close proximity to metallic silver particles. Time-resolved measurements showed that the intensity increase was due in part to a silver-induced increase in the radiative decay rate. These results suggest the use of labeled antibodies conjugated to silver particles as ultrabright probes for imaging or analytical applications. |
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