Integrated petrographic, geochemical, and fluid inclusion study of fracture mineralization and associated host rock in selected Mississippian and Devonian carbonates extending from southeastern Alberta to northwestern British Columbia, Canada, aims to quantify the type and nature of fluid precipitated saddle dolomite and late calcite cement and their origin. Petrographic and isotopic evidence from both the Devonian and Mississippian fracture-filling carbonates indicate the presence of a hydrothermal fluid source. The δ18O isotopic values for the Devonian saddle dolomite (−14.62‰ to −3.75‰ VPDB, average −11.12‰) combined with enriched 87Sr/86Sr isotopic ratios (0.70827–0.71599, average 0.71006) and higher homogenization temperatures (Th = 74–194.6 °C, average 126.8 °C) and salinity values (7.7–26.6 wt.% NaCl, average 16.2 wt.% NaCl) show significant differences from the Mississippian saddle dolomite, which is characterized by less negative δ18O isotopic values (−12.53‰ to −7.82‰ VPDB, average −9.14‰), less radiogenic 87Sr/86Sr isotopic ratios (0.70859–0.70943, average 0.70887), and lower homogenization temperatures (Th) and salinity values of fluid inclusions (87.6–214.2 °C, average 136.3 °C; 2.0–13.2 wt.% NaCl, average 9.6 wt.% NaCl). Later fracture- and vug-rimming blocky calcite cement records comparable or slightly lower values of δ18O (−16.31‰ to −4.08‰ VPDB, average −9.76‰) and 87Sr/86Sr (0.70784–0.709743, average 0.70868) and much lower salinity values (0–22.5 wt.% NaCl, average 2.86 wt.% NaCl) for samples mostly from the Mississippian age group. These results possibly suggest two different hydrothermal episodes related to early (Antler) and late (Laramide) tectonic events that affected the Western Canada Sedimentary Basin with possible compartmentalization of hydrothermal systems and their associated brines in the basin.