The St Louis River (SLR) is the second largest tributary to Lake Superior and a designated area of concern (AOC) due to past industrial contamination of organic chemicals and heavy metals, including mercury (Hg). Sediments, prey items (odonates, mixed benthic invertebrates, and spiders), and game fish were targeted within this study to determine if industrial sources of Hg were bioaccumulating into the food web. Samples from SLR were directly compared to a reference site in the Bad River, WI to assess if legacy contamination resulted in elevated Hg concentrations in comparisons to background regions with no known point sources. Carbon and nitrogen isotopes were also employed as food web tracers to determine if dietary shifts within the estuary or migration influenced Hg source signatures. These data are a part of a larger study and associated journal article documented in Science of the Total Environment (https://doi.org/10.1016/j.scitotenv.2021.146284).