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(F-F0)=(F-Fmax)/[1+(KD/[protein])^n] Here, F and F0 are the fluorescence intensities of the drugs in the presence and absence of protein, respectively, Fmax is the maximum fluorescence intensity of protein-drug complex, KD is the equilibrium dissociation coefficient that quantifies the strength of the protein-drug interaction, and n is the Hill coefficient, which is generally regarded as a measure of the association cooperativity. Plot F versus [protein], and fit the plots with the above function. Then you will get the value of KD, n and Fmax. The binding constant is actually the reciprocal of KD. ref: J. Chem. Soc., Faraday Trans. 1998, 94, 2083-2087 Nat. Nanotechnol. 2009, 4, 577-580 |
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