Deep Springs Valley (DSV) is a hydrologically isolated valley between the White and Inyo mountains that is commonly excluded from regional paleohydrology and paleoclimatology. Previous studies showed that uplift of Deep Springs ridge (informal name) by the Deep Springs fault defeated streams crossing DSV and hydrologically isolated the valley sometime after eruption of the Pleistocene Bishop Tuff (0.772 Ma). Here, we present tephrochronology and clast counts that reaffirms interruption of the Pliocene–Pleistocene hydrology and formation of DSV during the Pleistocene. Paleontology and infrared stimulated luminescence (IRSL) dates indicate a freshwater lake inundated Deep Springs Valley from ca. 83–61 ka or during Late Pleistocene Marine Isotope Stages 5a (MIS 5a; ca. 82 ka peak) and 4 (MIS 4; ca. 71–57 ka). The age of pluvial Deep Springs Lake coincides with pluvial lakes in Owens Valley and Columbus Salt Marsh and documents greater effective precipitation in southwestern North America during MIS 5a and MIS 4. In addition, we hypothesize that Deep Springs Lake was a balanced-fill lake that overflowed into Eureka Valley via the Soldier Pass wind gap during MIS 5a and MIS 4. DSV hydrology has implications for dispersal and endemism of the Deep Springs black toad (Anaxyrus exsul).