Despite consistent findings associating autonomic activity, such as resting heart rate, with antisocial behavior, the research connecting autonomic variables to related phenotypes, such as psychopathy and sensation seeking, has been mixed. The existing research in this area has been limited by underpowered samples, focused predominantly on incarcerated males, frequently dichotomized samples into “psychopaths” and controls, and failed to consider potential gender differences. The current study sought to address some of these limitations using a relatively large undergraduate sample (N = 453), four measures of autonomic activity (e.g., resting heart rate, resting skin conductance, heart rate reactivity, and skin conductance reactivity), a sensation seeking scale, and two measures of psychopathic traits. In order to thoroughly assess possible gender differences, the analyses were conducted for males and females separately. Few significant associations were found between the autonomic and psychological variables, and most became insignificant after controlling for age and race and correcting for multiple comparisons. The current study offers little support for an association between autonomic activity and sensation seeking or psychopathic traits.