Nocturnal car traffic often results in amphibian casualties, especially during rainy nights. The behavior of amphibians presumably influences their vulnerability to mortality on the road, but this hypothesis remains untested. We investigated the behavioral response of individuals of six species of amphibians on roads when confronted by an approaching vehicle. We first conducted a field study consisting of 50 night-driving surveys over 4 yr during which we recorded the behavior (i.e., moving or immobile) of frogs, toads, tree frogs, and salamanders encountered on a 20-km stretch of road. In an effort to tease apart the effects of headlights and the sound of motors on amphibian behavior, we carried out a field experiment on a test road where we exposed individuals to different car-associated stimuli. Here, we tested the hypothesis that simultaneous exposure to headlights and the sound of a car motor would elicit a stronger response than exposure to a single stimulus or a control. Based on the observations of the 2767 individuals in the field survey, immobility was the most common response to the approach of a car (mean probability of 0.82 of remaining immobile); the response differed across species but depended on the season of the survey (May–June vs. July–September). Similarly, the 91 individuals included in the field experiment were more likely to move during the control treatment than during any of the car-associated treatments. The combined stimuli elicited the strongest response, followed by the headlights-only and the motor-only treatments. Spring peepers (Pseudacris crucifer) tended to move more often than the other species we tested in the field experiment, which suggests they spend less time on the road and are less vulnerable to traffic mortality than other species. Both the field survey and field experiment consistently indicated that amphibians tend to remain immobile at the approach of a vehicle. This behavior highlights the vulnerability of amphibians to road traffic and should be considered in measures to mitigate road impacts.