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Equilibrium calcium binding of calbindin-AFF variants. (A) Binding of calcium to Q65′ (●) and (Q65′+W66) (○), detected by pyrene excimer fluorescence. (B) Binding of calcium to Q65′ (●), (Q65′+W66) (○), and (Q65′+W66′) (gray triangles) monitored by BODIPY fluorescence. Lines are best fits of the data to the Hill equation.
