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    Development of a quantitative fluorescence-based ligand-binding assay


    Breen, Conor and Raverdeau, Mathilde and Voorheis, H. Paul (2016) Development of a quantitative fluorescence-based ligand-binding assay. Scientific Reports, 6 (25769). ISSN 2045-2322

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    Abstract

    A major goal of biology is to develop a quantitative ligand-binding assay that does not involve the use of radioactivity. Existing fluorescence-based assays have a serious drawback due to fluorescence quenching that accompanies the binding of fluorescently-labeled ligands to their receptors. This limitation of existing fluorescence-based assays prevents the number of cellular receptors under investigation from being accurately measured. We have developed a method where FITC-labeled proteins bound to a cell surface are proteolyzed extensively to eliminate fluorescence quenching and then the fluorescence of the resulting sample is compared to that of a known concentration of the proteolyzed FITC-protein employed. This step enables the number of cellular receptors to be measured quantitatively. We expect that this method will provide researchers with a viable alternative to the use of radioactivity in ligand binding assays.

    Item Type: Article
    Additional Information: This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ Cite as: J. Breen, Conor & Raverdeau, Mathilde & Voorheis, H. Paul. (2016). Development of a quantitative fluorescence-based ligand-binding assay. Scientific Reports. 6. 25769. 10.1038/srep25769.
    Keywords: quantitative fluorescence-based ligand-binding assay;
    Academic Unit: Faculty of Science and Engineering > Biology
    Item ID: 10986
    Identification Number: https://doi.org/10.1038/srep25769
    Depositing User: Conor Breen
    Date Deposited: 21 Aug 2019 14:47
    Journal or Publication Title: Scientific Reports
    Publisher: Nature Publishing Group
    Refereed: Yes
    URI:

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