This study introduces a novel muscle activation analysis based on surface electromyography (sEMG) signals to assess the muscle's after-fatigue condition. Previous studies have mainly focused on the before-fatigue and fatigue conditions. However, a comprehensive analysis of the after-fatigue condition has been overlooked. The proposed method analyzes muscle fatigue indicators at various maximal voluntary contraction (MVC) levels to compare the before-fatigue, fatigue, and after-fatigue conditions using amplitude-based, spectral-based, and muscle fiber conduction velocity (CV) parameters. In addition, the contraction time of each MVC level is also analyzed with the same indicators. The results show that in the after-fatigue condition, the muscle activation changes significantly in the ways such as higher CV, power spectral density shifting to the right, and longer contraction time until exhaustion compared to the before-fatigue and fatigue conditions. The results can provide a comprehensive and objective evaluation of muscle fatigue and recovery, which can be helpful in clinical diagnosis, rehabilitation, and sports performance.