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Updated: Aug 17, 2026

Fluorescence detection methods for microfluidic droplet platforms
Published on: December 10, 2011
Analysis of fluorescence quenching data by the method of Levine
Abstract:
The quenching of intrinsic protein fluorescence upon binding of ligand is a relatively quick and easy method for studying these interactions. The method of Steiner et al. (J. Biol. Chem. 241: 560, 1966) and the method of Levine (Clin. Chem. 23: 2292, 1977) have been used by different authors to interpret the data obtained from fluorescence-quenching studies. However, as is shown here, the method of Levine is not strictly valid when the protein contains more than one binding site for the ligand. Therefore, this method only approximates binding constants from such data, with an accuracy that depends on the specific binding properties of the interactants.
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