This article presents the results of an empirical study looking at the relationships among perceived organizational support, anticipated and actual resource loss perceptions, and three dimensions of psychological distress, i.e., depression, anxiety, and fatigue. Moreover, the trait of negative affectivity was used as a moderator of the effects of perceived organizational support. We used conservation of resources theory as a framework for the study. Essentially, conservation of resources theory states that individuals strive to retain resources so as to build their well-being and that stress symptoms appear when individuals perceive or anticipate resource loss and/or think they won’t be able to gain resources (Hobfoll, 2002). Our reasoning in this paper is that perceived organizational support reduces employees’ perception of resource loss and of being unable to gain resources, and through that psychological mechanism reduces psychological distress. We also theorized that these mechanisms would be stronger among individuals with high levels of negative affectivity, as these individuals may be more fragile and would particularly need to be supported in order for them to build their resilience. We conducted an empirical study testing the above predictions using a sample of 138 contact service employees from four Canadian organizations. Consistent with predictions, analyses revealed that perceived organizational support was negatively related to perceived actual resource loss (yet was unrelated to anticipated resource loss). Moreover, perceived organizational support acted as a reducer of depression, anxiety, and fatigue, through the mediation of a reduced perception of actual resource loss. Moreover, perceived organizational support’s effects were amplified under conditions of high trait negative affectivity. This study’s results (a) contribute to improve our understanding of the dynamics of personal and social resources in the workplace, (b) highlight the importance of resource loss perceptions in the stress process, and (c) clarify the process through which perceived organizational support influences employees’ psychological well-being. This study’s findings also have implications for practice as they suggest that organizations can reduce employee distress either by demonstrating indications of support to employees or by encouraging the development of more positive perceptions of personal resources. Finally, we also discuss the meaning of the present findings for how conservation of resources theory principles can be integrated within the theory of perceived organizational support.