The study explores the nexus between ambidextrous leadership and ambidextrous innovation (exploratory and exploitative innovation). The mediating role of psychological safety and job autonomy was assessed based on the leader-member exchange (LMX) and contingency theories. The researcher selected 684 private sector employees in Accra, Ghana. The research design employed in the study was time-lagged. Data on psychological safety and the components of ambidextrous innovation were collected at T1, and data on ambidextrous leadership and job autonomy were collected at T2. The responses were analyzed using JASP software, applying Bootstrap resampling with 10,000 replications. The results indicate that psychological safety and job autonomy mediate the significant positive relationship between ambidextrous leadership and the two components of ambidextrous innovation. The present study exclusively focused on private-sector employees. Future research should also include public sector employees to help generalize the results. To encourage ambidextrous innovation, management should cultivate an environmental context where employees feel safe to take interpersonal risks and have autonomy in decision-making related to their job responsibilities. This study undertook a comprehensive examination of the two components of innovation, an aspect that previous studies have often overlooked. Moreover, the study expands the scope by considering psychological safety and job autonomy as mediating factors in the relationship under investigation.