Electromyographic (EMG) amplitude and mechanical tension are directly related during isometric contraction. Maximal voluntary isometric contractions are typically elicited through two different procedures; resisting a load, which is eccentric in nature, and contracting against an immovable object, which is concentric in nature. A wealth of literature exists indicating that EMG amplitude during concentric contractions is greater than that of eccentric contractions of the same magnitude. However, the effects of different methods to elicit isometric contraction on EMG amplitude have yet to be investigated. The purpose of this study was to compare EMG amplitudes under different loading configurations designed to elicit isometric muscle contraction. Twenty healthy volunteers (10 males and 10 females, age = 23 ± 2 yrs, height = 1.7 ± 0.09 m, mass = 69.9 + 16.8 kg) performed a maximal voluntary plantarflexion effort for which the vertical ground reaction force (GRFv) sampled from a force plate and surface EMG of the soleus were recorded. Participants then performed isometric plantarflexion at 20%, 30%, 40%, and 50% GRFv max in a seated position, from a neutral ankle position, under two different counterbalanced isometric loading conditions (concentric and eccentric). For concentric loading conditions, the subject contracted against an immovable resistance to the specified %GRFv identified via visual and auditory feedback. For eccentric loading conditions, subjects contracted against an applied load placed on the distal anterior thigh that produced the specified %GRFv. This applied load had the tendency to force the ankle into dorsiflexion. Therefore, plantarflexion force, in an attempt to maintain the ankle in a neutral position, resisted lengthening of the plantarflexor musculature, thus representing eccentric loading during an isometric contraction. Mean EMG amplitude was compared across loading levels and types using a 2 (loading type: concentric, eccentric) × 4 (loading level: 20%, 30%, 40%, 50% GRFv) repeated-measures ANOVA. The main effect for loading level was significant ( p = 0.007). However, the main effect for loading type, and the loading type × loading level interaction were non-significant ( p > 0.05). The present findings provide evidence that isometric muscle contractions loaded in either concentric or eccentric manners elicit similar EMG amplitudes, and are therefore comparable in research settings.