Distinguishing gastrocnemius and soleus muscle function is relevant for treating gait disorders in which abnormal plantarflexor activity may contribute to pathological movement patterns. Our objective was to use experimental and computational analysis to determine the influence of gastrocnemius and soleus activity on lower limb movement, and determine if anatomical variability of the gastrocnemius affected its function. Our hypothesis was that these muscles exhibit distinct functions, with the gastrocnemius inducing limb flexion and the soleus inducing limb extension. To test this hypothesis, the gastrocnemius or soleus of twenty healthy participants was electrically stimulated for brief periods (90 ms) during mid-or terminal stance of a random gait cycle. Muscle function was characterized by the induced change in sagittal pelvis, hip, knee, and ankle angles occurring during the 200 ms after stimulation onset. Results were corroborated with computational forward dynamic gait models, by perturbing gastrocnemius or soleus activity during similar portions of the gait cycle. Mid- and terminal stance gastrocnemius stimulation induced posterior pelvic tilt, hip flexion and knee flexion. Mid-stance gastrocnemius stimulation also induced ankle dorsiflexion. In contrast, mid-stance soleus stimulation induced anterior pelvic tilt, knee extension and plantarflexion, while late-stance soleus stimulation induced relatively little change in motion. Model predictions of induced hip, knee, and ankle motion were generally in the same direction as the experiments, though the gastrocnemius’ results were shown to be quite sensitive to its knee-to-ankle moment arm ratio.