Electromyographic (EMG) raw signals are sensitive to intrinsic and extrinsic factors. Consequently, EMG normalization is required to draw proper interpretations of standardized data. Specific recommendations are needed regarding a relevant EMG normalization method for participants who show atypical EMG patterns, such as post-stroke subjects. This study compared three EMG normalization methods ("isometric MVC", "isokinetic MVC", "isokinetic MVC kinematic-related") on muscle activations and the antagonist-agonist co-contraction index. Fifteen post-stroke subjects and fifteen healthy controls performed active elbow extensions, followed by isometric and isokinetic maximum voluntary contractions (MVC). Muscle activations were obtained by normalizing EMG envelopes during active movement using a reference value determined for each EMG normalization method. The results showed no significant difference between the three EMG normalization methods in post-stroke subjects on muscle activation and the antagonist-agonist co-contraction index. We highlighted that the antagonist-agonist co-contraction index could underestimate the antagonist co-contraction in the presence of atypical EMG patterns. Based on its practicality and feasibility, we recommend the use of isometric MVC as a relevant procedure for EMG normalization in post-stroke subjects. We suggest combined analysis of the antagonist-agonist co-contraction index and agonist and antagonist activations to properly investigate antagonist co-contraction in the presence of atypical EMG patterns during movement.