In biochemistry, an Eadie-Hofstee diagram (also Woolf-Eadie-Augustinsson-Hofstee or Eadie-Augustinsson plot) is a graphical representation of enzyme kinetics in which reaction velocity is plotted as a function or the velocity vs. substrate concentration ratio:

where v represents reaction velocity, K_m is the Michaelis-Menten constant, [S] is the substrate concentration, and v_max is the maximum reaction velocity.

Like other techniques that linearize the Michaelis-Menten equation, the Eadie-Hofstee plot allows for rapid identification of important kinetic terms like K_m and v_max. It is also more robust against error-prone data than the Lineweaver-Burke plot, particularly because it gives equal weight to data points in any range of substrate concentration or reaction velocity. (The Lineweaver-Burke plot unevenly weights such points.)

One drawback from the Eadie-Hofstee approach is that neither ordinate or abscissa represent independent variables: both are dependent on reaction velocity. Thus any experimental error will be present in both axes.