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Diffusion and binding properties investigated by Fluorescence Correlation Spectroscopy (FCS).

Grünwald, D. and Cardoso, M. Cristina and Leonhardt, H. and Buschmann, V. (2005):
Diffusion and binding properties investigated by Fluorescence Correlation Spectroscopy (FCS).
In: Current pharmaceutical biotechnology, pp. 381-386, 6, (5), ISSN 1389-2010, [Online-Edition: http://www.cardoso-lab.org/publications/Grunwald_2005.pdf],
[Article]

Abstract

During the last years, Fluorescence Correlation Spectroscopy (FCS) has proven to be a powerful tool for basic research in many applications. The combination of a minimal detection volume in the femtoliter range coupled with very high sensitivity extends the possibilities to design sensitive homogeneous tests. In this article we illustrate the analysis of binding processes with FCS based on the changes in diffusion characteristics of GFP upon binding to an antibody. Problems induced by highly heterogeneous samples are discussed and differences of GFP binding to a monoclonal and a polyclonal antibody are shown and analyzed. We stress data processing, limitations and useful approximations in FCS methodology. Basic ideas of data acquisition and processing as well as new developments and applications are presented.

Item Type: Article
Erschienen: 2005
Creators: Grünwald, D. and Cardoso, M. Cristina and Leonhardt, H. and Buschmann, V.
Title: Diffusion and binding properties investigated by Fluorescence Correlation Spectroscopy (FCS).
Language: English
Abstract:

During the last years, Fluorescence Correlation Spectroscopy (FCS) has proven to be a powerful tool for basic research in many applications. The combination of a minimal detection volume in the femtoliter range coupled with very high sensitivity extends the possibilities to design sensitive homogeneous tests. In this article we illustrate the analysis of binding processes with FCS based on the changes in diffusion characteristics of GFP upon binding to an antibody. Problems induced by highly heterogeneous samples are discussed and differences of GFP binding to a monoclonal and a polyclonal antibody are shown and analyzed. We stress data processing, limitations and useful approximations in FCS methodology. Basic ideas of data acquisition and processing as well as new developments and applications are presented.

Journal or Publication Title: Current pharmaceutical biotechnology
Volume: 6
Number: 5
Divisions: 10 Department of Biology
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10 Department of Biology > Cell Biology and Epigenetics
Date Deposited: 06 Mar 2010 07:51
Official URL: http://www.cardoso-lab.org/publications/Grunwald_2005.pdf
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