Maternal care is considered a universal and even cross-species set of typical behaviors, which are necessary to determine the social development of children. In humans, most research on mother–infant bonding is based on Western cultures and conducted in European and American countries. Thus, it is still unknown which aspects of mother–infant behaviors are universal and which vary with culture. Here we test whether typical mother–infant behaviors of affect-communication and affect-regulation are equally represented during spontaneous interaction in Palestinian-Arab and Jewish cultures. 30 Palestinian-Arab and 43 Jewish mother–infant dyads were recruited and videotaped. Using AffectRegulation Coding System (ARCS), we behaviorally analyzed the second-by-second display of valence and arousal in each participant and calculated the dynamic patterns of affect co-regulation. The results show that Palestinian-Arab infants express more positive valence than Jewish infants and that Palestinian-Arab mothers express higher arousal compared to Jewish mothers. Moreover, we found culturally-distinct strategies to regulate the infant: increased arousal in Palestinian-Arab dyads and increased mutual affective match in Jewish dyads. Such cross-cultural differences in affect indicate that basic features of emotion that are often considered universal are differentially represented in different cultures. Affect communication and regulation patterns can be transmitted across generations in early-life socialization with caregivers.