Prosocial safety behavior (PSB) is a critical element of workforce participation in the promotion of safety in the workplace. This study aims to examine the influence of various antecedents on PSB. Based on an analysis of the existing literature on prosocial behaviors, we investigate whether emotional intelligence (EI) and perceived organizational support (POS) significantly influence PSB. Furthermore, we analyze the mediating role of employees’ intrinsic motivation (IM) and affective commitment (AC) in these relationships. The research was conducted by administering an online questionnaire to a large sample of 488 workers employed in safety-critical industries based in the United States and the United Kingdom with the support of an online platform (N = 346; N = 142). Statistical analyses, conducted using a multi-group approach, revealed that EI had a direct effect on PSB in the two samples analyzed. In the US sample, we found that IM mediated the influence of EI and POS on PSB. In the UK sample, none of the mediation hypotheses were supported. The study provides insights into the individual and organizational factors that promote the development of a prosocial orientation in managing workplace safety issues, revealing the importance of recognizing the role of EI as a significant person-related antecedent supporting PSB. The statistical evidence from the study suggests that organizations that desire to facilitate the expression of PSB in their workforce may consider investing in training programs to enhance the EI of their employees and designing work conditions that facilitate high levels of intrinsic motivation.