Spatio-temporal partitioning is a viable mechanism for minimizing resource competition among sympatric species. The occurrence of sympatric large carnivores – tiger Panthera tigris, leopard Panthera pardus and dhole Cuon alpinus – in forests of the Indian subcontinent is complemented with high dietary overlap. We characterized temporal and spatial patterns of large carnivores with major prey species using photo-captures from 50 camera trap stations in Mudumalai Tiger Reserve, Western Ghats during 2008–2010. We tested whether major prey species' activity and spatial use acted as drivers for coexistence among large carnivores. Tiger exhibited cathemeral activity in the night and is spatially correlated with sambar and gaur, supporting hypotheses related to large-sized prey. Leopard was active throughout the day and is spatially correlated with almost all prey species with no active separation from tiger. Dhole exhibited diurnal activity and spatial use in relation to chital and avoided felids to a certain extent. Leopard exhibited spatial correlation with tiger and dhole, while tiger did not correlate with dhole. Leopard exhibited relatively broader temporal and spatial tolerance due to its generalist nature, which permits opportunistic exploitation of resources. This supports the hypothesis that predators actively used areas at the same time as their principal prey species depending upon their body size and morphological adaptation. We conclude that resource partitioning in large carnivores by activity and spatial use of their principal prey governs spatio-temporal separation in large carnivores.