To measure perceived interdependence, we designed a scale that explicitly measures how individuals’ feelings and outcomes covary with the outcomes of specific partners (e.g., “When [target] succeeds, I feel good.”, “[target] and I rise and fall together”). This new scale relies on the concept of fitness interdependence, i.e., the degree to which two or more organisms influence each other’s success in replicating their genes. Based on two studies of MTurk participants (N1 = 198, N2 = 216), we confirm that the scale has good overall reliability (Cronbach’s alpha = 0.81 to 0.91; McDonald’s omega = 0.89 to 0.96) as well as an underlying two-factor structure distinguishing the perceived outcome interdependence with the target and the emotional affect towards a target’s outcomes. Our scale also predicted participants’ willingness to help interdependent others. Studies 3 and 4 (N3 = 695, N4 = 629) document the validity and reliability of the scale with additional measures in new samples. The results from these studies demonstrate that the perceived interdependence scale has good discriminant, convergent, and concurrent validity as well as test-retest reliability across a 14 day period. Taken together, our results suggest that perceived interdependence offers a useful framework for future work on cooperation and social behavior. However, there are still many open questions about the cognitive architecture underlying perceptions of interdependence and how perceived interdependence interacts with genetic relatedness.