Unique to South-east Asia, Lao People's Democratic Republic contains extensive habitat for tigers and their prey within a multiple-use protected area system covering 13% of the country. Although human population density is the lowest in the region, the impact of human occurrence in protected areas on tiger Panthera tigris and prey populations was unknown. We examined the effects of human–carnivore conflict on tiger and prey abundance and distribution in the Nam Et-Phou Louey National Protected Area on the Lao–Vietnam border. We conducted intensive camera-trap sampling of large carnivores and prey at varying levels of human population and monitored carnivore depredation of livestock across the protected area. The relative abundance of large ungulates was low throughout whereas that of small prey was significantly higher where human density was lower. The estimated tiger density for the sample area ranged from 0.2 to 0.7 per 100 km2. Tiger abundance was significantly lower where human population and disturbance were greater. Three factors, commercial poaching associated with livestock grazing followed by prey depletion and competition between large carnivores, are likely responsible for tiger abundance and distribution. Maintaining tigers in the country's protected areas will be dependent on the spatial separation of large carnivores and humans by modifying livestock husbandry practices and enforcing zoning.