Two research traditions explain the way we deal with emotional situations: emotional intelligence (EI) and emotion regulation (ER). EI refers to the individual differences in the knowledge, identification, and regulation of emotions. ER describes processes in which emotions are experienced, expressed, and altered. Our study examined the EI-ER link and their moderating role on affective responses. We used self-report questionnaires and a cognitive reappraisal (CR) task, in which subjective affective responses were registered. We found that higher levels of ER difficulties correlated with lower EI. Gender had an overall impact on affective changes, indicating a more unpleasant and more arousing affective state for women compared with men. Regarding the moderating role of EI and ER difficulties, the ability to utilize emotions (Utilization) decreased the valence into a more unpleasant direction, similar to the effect of the inability to identify and differentiate emotions (Clarity). A weak control over emotions (Impulse), however, increased the valence into a more pleasant direction. The lack of attention to emotional signals (Awareness) marginally decreased the initial intensity (i.e., lower level of arousal). We demonstrated that EI and ER have distinctive routes and a different influence on the affective outcome defined by valence and arousal ratings: (1) EI has an impact through the utilization of emotions mainly on the valence dimension; and (2) individual differences in ER have a moderating effect on both valence and arousal dimensions. This study provided evidence on how individual differences contribute to a successful ER process when using a CR strategy.