Objectives: Gendered inequalities in workplace stress are linked to sex-specific health trajectories that are poorly understood. Measuring gendered inequalities is challenging but necessary to better explain individual differences in occupational health. The aim of this exploratory, retrospective study was to create a measure of occupational gender-roles and use structural equation models to investigate pathways linking layers of gendered factors to workplace stress, allostatic load, and mental health in a sample of psychiatric hospital workers (N = 192). Methods: Individual-level gender-roles were measured with the Bem Sex-Role Inventory Short-Form. Occupational gender-roles were measured using a novel web-based survey approach. Sex-specific allostatic load indices were constructed using 23 biomarkers (e.g., neuroendocrine, immune, cardiovascular, and metabolic). Workplace stress was assessed using the Job Content Questionnaire and the Effort-Reward at Work Questionnaire. Depressive symptoms were assessed with the Beck Depression Inventory-II, burnout symptoms with the Maslach Burnout Inventory – General Survey, and trauma symptoms with the PTSD Civilian Checklist. Results: Individual-level masculine gender-roles were positively associated with psychological demands (R² = 0.103) and social support (R² = 0.078). Masculine and feminine occupational gender-roles were positively associated with decisional latitude (R² = 0.157) and effort-reward ratio (R² = 0.058). Both individual masculine and feminine gender-roles had protective effects on depressive symptoms (R² = 0.289) and burnout symptoms (R² = 0.306) but only individual masculine gender-roles had protective effects on trauma symptoms (R² = 0.198). We found no association between occupational gender-roles and mental health and allostatic load. Conclusion: Beyond individual gender-roles, our study shows the utility of measuring occupational gender-roles to delineate associations between workplace stressors and mental health that should be applied in future studies of sex differences in occupational health.