Although job stress has become an issue of great concern over the last decades both internationally and nationally, there still remains a paucity of research in the Ghanaian insurance industry. This study therefore examined the relationship between antecedent variables (work overload, role conflict and role ambiguity) and employee job stress in the insurance industry in Ghana. Using a descriptive cross-sectional design with a survey questionnaire, 212 employees were selected to participate in the study. Pearson correlation and a two-step hierarchical regression were used to test the proposed hypotheses. Results of the analysis revealed that work overload and role conflict rather than role ambiguity were positively related to job stress. Implications for theory and practice are later discussed in the study.