Measurement of Complex I Functionality
The functionality of complex I is measured by the Iberius by first treating the cells with oligomycin to inhibit the ATP synthase and decrease the electron flux bringing complex I closer to equilibrium (Figure 3).
The cells are then treated with multiple small doses of an uncoupler to increase the electron flux and drive complex I out of equilibrium.
The flux is then plotted as a function of ΔGI and the gradient of the response close to equilibrium calculated from a linear regression.