The study examined attitudes toward work–life balance (WLB) using survey data from 292 employees in an Australian university. Organisational culture, as it relates to how and when employees can use WLB initiatives, was investigated. In particular, the study examined employees' differing perceptions of organisational culture within a single organisation, with particular reference to: managerial support of WLB initiatives, career consequences of using WLB initiatives, organisational time expectations that may interfere with non-work activities, and the level of employee control over workload and when employees can take time off. As predicted, perceptions of managerial support of WLB initiatives were related to initiative use; however, overall initiative use was not related to perceived career consequences (e.g. using initiatives will damage one's career progress), organisational time expectations (e.g., long working hours), or levels of employee control (e.g., employee control over workload). Results showed that perceptions of a supportive organisational culture were positively related to organisational commitment and negatively related to work–life conflict and employees' intentions to leave. Further, the study results show that employees' perception of organisational culture was more strongly associated with the outcomes (e.g., work attitudes, work–life conflict) than the number of WLB initiatives used by employees.