To construct an Eadie-Hofstee plot, follow these steps:
Measure the initial reaction velocity (v) at various substrate concentrations ([S]). Calculate v/[S] for each data point. Plot v on the y-axis and v/[S] on the x-axis.
The resulting graph will be a straight line with a slope of -Km and an intercept on the y-axis equal to Vmax. The x-intercept of the plot gives Vmax/Km.
