Our various daily activities continually require regulation of our internal state. These regulatory processes covary with changes in High Frequency Heart Rate Variability (HF-HRV), a marker of parasympathetic activity. Specifically, incidental increases in HF-HRV accompany positive social engagement behavior and prosocial action. Little is known about deliberate regulation of HF-HRV and the role of voluntary parasympathetic regulation in prosocial behavior. Here, we present a novel biofeedback task that measures the ability to deliberately increase HF-HRV. In two large samples, we find that a) participants are able to voluntarily upregulate HF-HRV, and b) variation in this ability predicts individual differences in altruistic prosocial behavior, but not non-altruistic forms of prosociality, assessed through 14 different measures. Our findings suggest that self-induction of parasympathetic states is involved in altruistic action. The biofeedback task may provide a measure of deliberate parasympathetic regulation, with implications for the study of attention, emotion, and social behavior.