Snow-dominated and glacierized catchments are important sources of fresh water for biological communities and for populations living in mountain valleys. Gaining a better understanding of the runoff origin and of the hydrological interactions between meltwater, streamflow and groundwater is critical for natural risk assessment and mitigation as well as for effective water resource management in mountain regions. This study is based on the use of stable isotopes of water and electrical conductivity as tracers to identify the water sources for runoff and groundwater and their seasonal variability in a glacierized catchment in the Italian Alps. Samples were collected from rainfall, snow, snowmelt, ice melt, spring and stream water (from the main stream at different locations and from selected tributaries) in 2011, 2012 and 2013. The tracer-based mixing analysis revealed that, overall, snowmelt and glacier melt were the most important end-members for stream runoff during late spring, summer and early fall. The temporal variability of the tracer concentration suggested that stream water was dominated by snowmelt at the beginning of the melting season (May–June), by a mixture of snowmelt and glacier melt during mid-summer (July–early August), and by glacier melt during the end of the summer (end of August–September). The same seasonal pattern observed in streamflow was also evident for groundwater, with the highest electrical conductivity and least negative isotopic values found during cold or relatively less warm periods, when the melt of snowpack and ice was limited. Particularly, the application of a two-component mixing model to data from different springs showed that the snowmelt contribution to groundwater recharge varied between 21% (±3%) and 93% (±1%) over the season, and the overall contribution during the three study years ranged between 58% (±24%) and 72% (±19%). These results provided new insights into the isotopic characterization of the study catchment presenting further understanding of the spatio-temporal variability of the main water sources contributing to runoff.