Introduction: The interpolated twitch technique is often used to assess voluntary activation (VA) of skeletal muscles. We investigated VA and the voluntary torque-superimposed torque relationship using either supramaximal nerve stimulation or better tolerated submaximal muscle stimulation, which is often used with patients. Methods: Thirteen healthy subjects performed maximal and submaximal isometric knee extensions with superimposed maximal or submaximal doublets (100 Hz). Results: Superimposed torque relative to potentiated resting doublets was smaller with maximal nerve than with submaximal muscle stimulation. Maximal VA was 87 ± 7% and 93 ± 5% for submaximal muscle and maximal nerve stimulation, respectively. The individual voluntary torque-superimposed torque relationships were more linear for submaximal muscle stimulation, possibly leading to less overestimation of VA. Conclusions: Submaximal muscle stimulation can be used to estimate VA in the knee extensors. It is less painful, and overestimation of VA may be less compared with maximal nerve stimulation. © 2012 Wiley Periodicals, Inc.