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  Home > JCE Print > Journal of Chemical Education > Issues > 2007  > August  >
In the Laboratory
Steady-State Fluorescence Anisotropy To Investigate Flavonoids Binding to Proteins
Christine M. Ingersoll and Christen M. Strollo
Department of Chemistry, Muhlenberg College, Allentown, PA 18104
Cover
August 2007
Vol. 84 No. 8
p. 1313

Abstract
A laboratory experiment using steady-state fluorescence polarization or anisotropy to determine the binding constant for a flavonoid–protein interaction is described. Using the intrinsic fluorescence of quercetin (a model flavonoid), the fluorescence anisotropy is measured as a function of human serum albumin (HSA) concentration. From the anisotropy data the fraction of bound quercetin at each protein concentration is calculated. Then, employing both linear (double reciprocal) and nonlinear regression analyses, the binding constant and Gibbs free energy change for the quercetin–HSA interaction are determined. This interdisciplinary exercise investigates a fluorescence technique not typically covered in the undergraduate curriculum and explores many aspects of chemistry such as equilibrium, thermodynamics, biochemical interactions, and data-analysis methods. The experiment can be completed in one, three-hour laboratory period and is appropriate for analytical or instrumental analysis, physical chemistry, and biochemistry courses.
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Instructions for students and notes for the instructor are available.
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Citation
Ingersoll, Christine M.; Strollo, Christen M. J. Chem. Educ. 2007, 84, 1313.
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Keywords
Analytical Chemistry; Fluorescence Spectroscopy; Food Science; Hands-On Learning / Manipulatives; Instrumental Methods; Laboratory Instruction; Phenols; Upper-Division Undergraduate
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History
Created:
Last Updated:
6/22/2007
7/5/2007
 Caution! 
Experiments, laboratory exercises, lecture demonstrations, and other descriptions of the use of chemicals, apparatus, instruments, computers, and computer interfaces are presented in the Journal of Chemical Education as illustrative of new or improved ideas or concepts in chemistry instruction and are directed at qualified teachers. Although every effort is made to assure and encourage safe practices and safe use of chemicals, the Journal of Chemical Education cannot assume responsibility for uses made of its published materials. Many chemicals are hazardous. Precautions for the safe use of hazardous chemicals and directions for their proper disposal are described in the Material Safety Data Sheets and on the labels. We strongly urge all those planning to use materials from our pages to make choices and to develop procedures for laboratory and classroom safety in accordance with local needs and situations.
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