This study investigates the modes of subsistence practices of a Middle Don Catacomb Culture community for which archaeological evidence suggests a reliance on seasonal mobility of humans and their herds. A sequential multi-isotope approach [stable carbon (δ13C), stable oxygen (δ18O), and strontium (87Sr/86Sr)] on tooth enamel of mainly domestic animals (cattle and sheep/goat) from the Middle Bronze Age settlement Ksizovo-1 provides new and important proxy data for the relationships between seasonal mobility and domestic animal husbandry in forest-steppe environments. Stable isotope analyses in the steppe zones of European Russia during the Bronze Age have so far mostly focused on human mobility and diet, with few exceptions. We combine the results of the isotope analysis with the archaeological evidence and use this information to draw conclusions about the organization of the pastoral lifestyles of the community and their herds. Incremental sampling of oxygen and carbon isotope values show a year-round intake of C3 plant forage and stay in the same ecology, and intra-tooth strontium isotopic variation almost completely corresponds with the local Sr range. The investigated birth periodicity shows different reproductive patterns of cattle and sheep/goat with signs of human interventions in the reproductive cycle. The results suggest small-scale animal movements limited to the regional ecology, which is comparable to the results of other investigations of Bronze Age husbandry practices in steppe environments using isotopic analysis. The study fosters a foundation based on empirical data for the understanding of pastoralist systems in the Eastern European Bronze Age aiming to move beyond the current reliance on hypotheses.