Abstract: In arid regions, groundwater is a vital resource that can also provide a long-term record of the regional water cycle. However, the use of groundwater as a paleoclimate proxy has been limited by the complex hydrology and the lack of appropriate chronometers. Applying 81Kr, a long-lived radioisotope tracer, we investigate the paleohydroclimate and subsurface water storage properties of the Nubian Sandstone Aquifer in the Negev Desert, Israel. Based on the spatial distributions of stable isotopes and the abundance of 81Kr, we resolve subsurface mixing and identify two distinct moisture sources of the recharge: one recent (<38 ky ago) from the Mediterranean and the other 361 ± 30 ky ago from the tropical Atlantic, both of which occurred under conditions of low orbital eccentricity comparable with that of the present. With this tracer, groundwater can now serve as a direct record of paleoprecipitation over land and of subsurface water storage from the mid-Pleistocene and onward.