Parenting behavior is a key factor in children's socio-emotional development. However, little is known about similarities and differences in maternal and paternal parenting behavior, as most studies have focused on mothers. The present study investigated similarities and differences in mothers' and fathers' parenting behavior during observed free play with their preschool children, in a Danish well-resourced sample. We examined differences in mean scores and associations between mothers' and fathers' sensitivity, intrusiveness and limit-setting assessed with the Coding Interactive Behavior instrument. Additionally, we conducted confirmatory factor analysis to test the model-fit between the measurement model and parental data. Exploratory factor analysis was conducted to investigate if maternal and paternal factor structures replicated the three parenting constructs, and to explore if certain parenting behaviors seemed specifically related to either mothering or fathering. Participants included 52 mothers, 41 fathers and their 5-year old children. Similar mean scores were found for mothers and fathers on all parenting constructs. Maternal and paternal parenting behavior were not correlated. Confirmatory factor analysis revealed a poor model-fit. For both mothers and fathers, latent factors related to sensitivity, intrusiveness and limit-setting emerged, which indicated that the Coding Interactive Behavior instrument was suitable for assessment of both maternal and paternal sensitivity, intrusiveness and limit-setting. However, item loadings suggested that the instrument assessed maternal sensitivity more accurately than paternal sensitivity in our sample. Two additional factors were retrieved for fathers, i.e. paternal performance and challenging behavior, and paternal teaching behavior. This finding may suggest that additional parenting constructs need to be developed for researchers to be able to thoroughly investigate similarities and differences in mothers' and fathers' parenting behavior. Despite difference in factor structure, we did not identify behaviors solely related to mothering or to fathering.