The aim of this study was to investigate the relationship between milk insulin-like growth factor-2 (IGF-2) concentration and milk yield in Anatolian water buffaloes. This study was conducted on milk samples from 80 Anatolian water buffaloes. Milk samples collected from buffaloes were evaluated for subclinical mastitis. For this purpose, the California mastitis test and somatic cell count were performed in milk samples taken from four different mammary lobes of buffaloes. The milk IGF-2 level was determined by an enzyme-linked immunosorbent assay kit. Milk IGF-2 concentration ranged from 21.6 ng/ml - 63.2 ng/ml in Anatolian water buffaloes. The IGF-2 concentration in Anatolian water buffaloes milk was 40.1±8.5 ng/ml. Milk IGF-2 was positively correlated with milk yield (r2=0.941, P0.01). Our results showed that milk IGF-2 concentration was associated with milk yield in Anatolian buffaloes. These findings show that locally synthesized IGF-2can affect milk yield. This study contributes to the understanding of the composition of buffalo milk, which has an important value in human nutrition. It is recommended to confirm the results of similar measurements in milk from other animal species used for human consumption.