The syn-tectonic breccia-hosted Mount Isa Cu deposit in northwest Queensland is the largest sediment-hosted Cu deposit in Australia. Whole-rock samples of chalcopyrite-rich Cu ore form an isochron with a Re–Os age of 1,372 ± 41 Ma. This age is more than 100 Ma younger than the previously accepted age of Cu ore formation, an Ar–Ar mineral age for biotite separated from the host rocks within the alteration envelope to the Cu orebody. This discrepancy cannot be unequivocally resolved due to a lack of other absolute geochronological constraints for Cu mineralisation or the deformation event associated with Cu emplacement. The 1,372 ± 41 Ma date may reflect (a) the time of Cu deposition, (b) the time of a hydrothermal event that reset the Re–Os signature of the Cu ore or (c) mixing of the Re–Os isotope systematics between the host rocks and Cu-bearing fluids. However, a range of published Ar–Ar and Rb–Sr dates for potassic alteration associated with Cu mineralisation also records an event between 1,350 and 1,400 Ma and these are consistent with the 1,372 Ma Re–Os age. The 1.8 Ga Eastern Creek Volcanics are a series of tholeiitic basalts with a primary magmatic Cu enrichment which occur adjacent to the Mount Isa Cu deposit. The whole-rock Os isotopic signature of the Eastern Creek Volcanics ranges from mantle-like values for the upper Pickwick Member, to more radiogenic/crustal values for the lower Cromwell Member. The Re–Os isotope signature of the Cu ores overlaps with those calculated for the two volcanic members at 1,372 Ma; hence, the Os isotope data are supportive of the concept that the Os in the Cu ores was sourced from the Eastern Creek Volcanics. By inference, it is therefore postulated that the Eastern Creek Volcanics are the source of Cu in the Mount Isa deposit, as both Os and Cu are readily transported by oxidised hydrothermal fluids, such as those that are thought to have formed the Cu orebody. The Pickwick Member yields a Re–Os isochron age of 1,833 ± 51 Ma, which is within error of previously reported age constraints. The initial ¹⁸⁷Os/¹⁸⁸Os isotopic ratio of 0.114 ± 0.067 (γOs = −0.7) is slightly subchondritic, and together with other trace element geochemical constraints, is consistent with a subcontinental lithospheric mantle source. The Pickwick Member records a minimum age of ca. 1.95 Ga for melt depletion in the subcontinental lithospheric mantle beneath the Mount Isa Inlier prior to the extraction of the magmas which formed the Eastern Creek Volcanics. This corresponds with the end of subduction-related magmatism along the eastern margin of the Northern Australian Craton, which included the Mount Isa Inlier.