The extent of ligand binding to a macromolecule is often measured spectroscopically. In the presence of a concentration [So] of the binding sites, the instrument detects the concentration of unbound ligand, [L], as a function of the (known) total ligand concentration, [Lo]. The latter is determined by adding ligand to a solution that does not contain the macromolecule so that the detection instrument responds to the total ligand concentration. The data are plotted as 1 -[L]/[Lo] versus [Lo] and compared to the theoretical model expressed by Eq (11.17). By combining Eqs (11.15) with (11.14), the quantity obtainable from experiment is expressed as:
which yields the fraction of ligand bounds to sites on the macromolecule:
Figure 11.9 is a plot of Eqs (11.20) and (11.17) for various values of K and [So] = 5 mM.
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