Adverse childhood experiences (ACE), such as emotional or physical abuse, can produce a lasting effect on the individual. The aim of this study was to investigate how ACE may impact electromyography (EMG) activity of the trapezius muscle in a novel experimental stress paradigm, in a sample of 120 healthy participants. The stress paradigm consisted of a memory task, in which participants were asked to memorize and recall as many words as possible, displayed on a screen. The study protocol included 2 identical experimental sessions (T 0 = 0 and T 1 = 6 months). EMG activity was analyzed using multilevel regression analysis. EMG activity was higher during the memory task compared to baseline, supporting the validity of the experimental EMG-stress paradigm. In addition, the EMG increase was attenuated during the second session. Analyses were indicative for a moderating effect of ACE on stress-induced EMG activity: higher ACE scores resulted in greater EMG reactivity. These associations were apparent for early ACE exposure (0–11 years) as well as for later exposure (12–17 years). The association between ACE and EMG reactivity remained significant but was much weaker at T 1 in comparison to T 0 , likely because of reduced unpredictability and uncertainty related to the experiment. In conclusion, this study showed that enduring liabilities occasioned by ACE in a non-clinical population can be studied using an experimental paradigm of EMG stress reactivity, contingent on the level of predictability of the stressor.