We determine whether mechanical asymmetries differ between dominant and non-dominant legs at fast treadmill speed. Stride temporal variables, derived from high-speed camera recordings, allowed to estimate leg and vertical stiffness through the sine-wave method in 31 uninjured males during treadmill running at 6.67 m.s−1. Lower limb dominance was determined by the triple-jump test. The asymmetry was expressed as dominant—non-dominant and indexed by the absolute asymmetry index (ASI). The lowest and highest mean ASI values were detected for contact time (1.69%) and flight time (5.66%), respectively; ASI values for spring-mass characteristics (2.6% ≤ leg and vertical stiffness, peak vertical force, change in vertical leg length and centre of mass vertical displacement ≤ 4.7%) were within this range. Inter-subject variability in ASI varied substantially among the seven analysed variables with larger and smaller range of variability in ASI found for flight time (0–16.56%) and contact time (0–3.47%), respectively. Because the magnitude of group mean ASI appears inconsistent among stride temporal and spring-mass characteristics, different biomechanical variables should not be used interchangeably to assess laterality effects at fast treadmill speed. The widespread ASI range also indicates that using a ‘fixed cut-off’ threshold is an arbitrary approach.