Increasing levels of human activity threaten wildlife populations through direct mortality, habitat degradation, and habitat fragmentation. Area closures can improve habitat quality for wildlife, but may be difficult to achieve where tourism or other economic drivers are a priority. Temporal closures that limit human use during specific times of day have potential to increase habitat quality for wildlife, while continuing to provide opportunities for human use. However, the effectiveness of daily temporal closures has not been tested. We assessed how implementation of a temporal road closure affected wildlife movements in Banff National Park. Parks Canada closed a popular 17 km stretch of road between 2000 and 0800 hours to improve habitat quality for wildlife. We assessed the effectiveness of the closure on nine mammal species using three sets of data: remote cameras, road surveys, and grizzly bear (Ursus arctos) GPS data. In all three analyses, wildlife detection rates on the road doubled during the closure while remaining unchanged in reference areas. Our strong and consistent results suggest temporal closures are an important conservation tool that can increase habitat quality for wildlife while minimizing effects on people.