During serpentinite rock weathering, Ni is concentrated in the regolith owing to residual and secondary enrichment, forming a Ni-rich deposit under tropical conditions. This study presents geochemical, isotopic, and mineralogical data for Ni from the Serra do Puma Complex (SPC) weathering profile in Carajas (Brazil). The Ni fluxes, redistribution, and Ni isotopic fractionation magnitudes were quantified along the entire weathering profile: rock, saprock, lower and upper saprolite, and limonite. The results show that chlorite and serpentine are the primary Ni-scavenging phases in saprolite, whereas Fe oxyhydroxides are the main Ni-hosting minerals in the limonite unit. The mass balance model confirmed a global Ni gain at the weathered profile scale, with larger Ni enrichment in the upper saprolite. The isotopic dataset obtained in this study contributes greatly to the knowledge of the current Ni cycle on regional and global scales. The detailed insights into Ni isotopes in the SPC, coupled with chemical and mineralogical composition, allow for the first time a Ni mass balance and Ni isotopic values for an entire weathering profile in an Amazonian context. The Ni isotopic profile agrees with the preferential retention of light Ni isotopes in the residual material relative to the parent rock during weathering processes, with the Δ60Ni limonite-saprock of −0.72‰. In the SPC, major isotopic fractionation was notably recorded during saprolitization, while limonitization was accompanied by an overall Ni loss without significant Ni isotopic fractionation. As already observed in previous studies, heavier Ni isotopes, preferentially leached during weathering processes, can be further mobilized downward and lost from the profile or incorporated in secondary minerals in the saprolite and limonite sections. The Ni isotopic mass balance model indicated that the Ni loss from the weathered serpentinite profile was isotopically heavy, in agreement with the isotopically heavier composition of the dissolved load of Amazonian rivers. The isotopically light Ni pool, associated with the significant Ni gain encountered in the upper saprolite, is notable in the SPC weathering profile and confirms the existence of a light Ni isotope reservoir in the continent.