We present the first Bayesian 14C modeling based on AMS ages from stratified sediments representing continuous occupation across the Early Bronze III/IV interface in the Southern Levant. This new high-precision modeling incorporates 12 calibrated AMS ages from Khirbat Iskandar Area C using OxCal 4.4.4 and the IntCal 20 calibration curve to specify the EB III/IV transition at or slightly before 2500 cal BCE. Our results contribute to the continuing emergence of a high chronology for the Levantine Early Bronze Age, which shifts the end of EB III 200–300 years earlier than the traditional time frame and increases the length of EB IV to about 500 years. Data from Khirbat Iskandar also help direct greater attention to the importance of sedentary communities through EB IV, in contrast to the traditional emphasis on non-sedentary pastoral encampments and cemeteries. Modeling of AMS data from Khirbat Iskandar bolsters the ongoing revision of Early Bronze Age Levantine chronology and its growing interpretive independence from Egyptian history and contributes particularly to re-examination of the EB III/IV nexus in the Southern Levant.