Few studies have assessed whether patterns of resource partitioning among sympatric marine predators vary between different habitats. This type of data is important for understanding food web functioning as well as for supporting an ecosystem-based fisheries management plan. In this study, we used δ13C and δ15N stable isotope analysis to assess the relative isotopic niche area (i.e., which species has the largest and smallest isotopic niche area within a study site) and core isotopic niche overlap between 299 sharks of 11 co-occurring shark species that vary in size, diet and behavior in three coastal study areas in South Florida. Overall, results show that the relative isotopic niche area and patterns of core isotopic niche overlap between some sympatric shark species varied between sites, suggesting plasticity in resource use and competitive interactions between shark species (e.g., 85% of blacktip shark Carcharhinus limbatus standard ellipse area overlapped with the blacknose shark C. acronotus ellipse in Biscayne Bay, but there was no overlap between these species’ ellipses in nearby Florida Bay). Therefore, patterns of resource use and trophic interactions documented among species from one site may not be applicable to a neighboring location. These findings may have implications for food web models that incorporate trophic data from different areas for a species when local data are unavailable.