['This study examined whether passive hamstring tissue stiffness and/or stretch tolerance explain the relationship between sex and hamstring extensibility.', 'Ninety healthy participants, 45 men and 45 women (mean ± SD; age 24.6 ± 5.9 years, height 1.72 ± 0.09 m, weight 74.6 ± 14.1 kg) volunteered for this study. The instrumented straight leg raise was used to determine hamstring extensibility and allow measurement of stiffness and stretch tolerance (visual analog pain score, VAS).', 'Hamstring extensibility was 9.9° greater in women compared to men (p = 0.003). VAS scores were 16 mm lower in women (p = 0.001). Maximal stiffness (maximal applied torque) was not different between men and women (p = 0.42). Passive stiffness (slope from 20-50° hip flexion) was 0.09 Nm.°(-1) lower in women (p = 0.025). For women, linear and stepwise regression showed that no predictor variables were associated with hamstring extensibility (adjusted r(2) = -0.03, p = 0.61). For men, 44% of the variance in hamstring extensibility was explained by VAS and maximal applied torque (adjusted r(2) = 0.44, p < 0.001), with 41% of the model accounted for by the relationship between higher VAS scores and lower extensibility (standardized β coefficient = -0.64, p < 0.001).'] Is lower hamstring extensibility in men compared to women explained by differences in stretch tolerance?