The open-air Acheulian site in Holon, Israel, was dated by the luminescence methods and by electron spin resonance (ESR). Situated in the coastal plain Quaternary Kurkar Group, the Holon site was first excavated in the late 1960s, when typical lower Paleolithic lithics and middle Pleistocene fauna were found. In order to date the site, new test pits were dug adjacent to the earlier excavations and the archaeological bed was exposed in a section comprising a series of paleosols and aeolianites. Alkali feldspars separated from the sediments were dated using the infrared stimulated luminescence and thermoluminescence signals, and quartz was dated using the optically stimulated luminescence signal. The age of the archaeological bed is constrained by two samples to 198,000 ± 22,000–201,000 ± 17,000 yr. The age of the base of the section is 240,000 ± 29,000 yr, and the age of the top is 81,000 ± 8000 yr. Two teeth from the archaeological bed, recovered from the original excavation collection, yielded an average ESR age of 204,000 ± 16,000 yr, calculated using the linear uptake model, which is in a very good agreement with the luminescence ages. These dates place Holon within the range of other late Acheulian and Acheulo-Yabrudian sites in this region such as Tabun E (younger chronology), Yabrud I (archaeological level 18), and Berekhat Ram.