The Cave of Pan is located on the N/NE slope of the hill of Oinoe (38°09′31.60′′N, 23°55′48.60′′E), west of modern Marathon. In rescue excavation campaigns during the last three years, among other finds, charcoal and seashell samples were also collected. The purpose of this study is the accelerator mass spectrometry (AMS) dating of the cave’s anthropogenic deposits and the calculation of the regional marine reservoir effect during the Neolithic period. For that purpose, 7 charcoal pieces and 1 seashell were dated. Our results show that the cave was used from the second quarter of the 6th millennium (Middle Neolithic period) until the beginning of the 5th millennium BC. Additionally, one sample collected from a depth of 2 cm from the present surface of the cave yielded an age falling within the 6th century AD, giving thus the absolute time span of the cave use. Moreover, the radiocarbon (14C) ages of one pair of charcoal-seashell samples showed that the marine reservoir age R(t) in the estuarine Marathon Bay region during the 5th millennium BC is 775±57 yr and the local sea surface reservoir deviation ΔR is found to be 402±63 14C yr (within 1σ).