Purpose This study examined gender differences in the association between work–family conflict and self-rated health and evaluated the effect of educational attainment. Method We used baseline data from ELSA-Brasil, a cohort study of civil servants from six Brazilian state capitals. Our samples included 12,017 active workers aged 34–72 years. Work–family conflict was measured by four indicators measuring effects of work on family, effects of family in work and lack of time for leisure and personal care. Results Women experienced more frequent work–family conflict, but in both genders, increased work–family conflict directly correlated with poorer self-rated health. Women’s educational level interacted with three work–family conflict indicators. For time-based effects of work on family, highly educated women had higher odds of suboptimal self-rated health (OR=1.54; 95 % CI=1.19–1.99) than less educated women (OR=1.14; 95 % CI=0.92–1.42). For strain-based effects of work on family, women with higher and lower education levels had OR=1.91 (95 % CI 1.48–2.47) and OR=1.40 (95 % CI 1.12–1.75), respectively. For lack of time for leisure and personal care, women with higher and lower education levels had OR=2.60 (95 % CI=1.95–3.47) and OR=1.11 (95 % CI=0.90–1.38), respectively. Conclusion Women’s education level affects the relationship between work–family conflict and self-rated health. The results may contribute to prevention activities.