The objective of the study was to establish correlation of seminal and serum IGF-1 with seminal attributes, estimate antioxidant potential of IGF-1 by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays and to study the effect of IGF-1 supplementation on semen cryopreservation. For this study, buffalo bulls were divided into sub-fertile (n = 2) and normal (n = 5) on the basis of sperm mass motility and individual motility. The serum IGF-1 concentration of normal bulls was greater than in sub-fertile bulls, but there was no difference in the seminal IGF-1 concentration among the groups. The values from correlation analyses indicated that serum IGF-1 concentration is positively correlated with semen mass motility and sperm concentration. In the second experiment, IGF-1 did not have antioxidant activities when assessed with DPPH and FRAP assays. In the third experiment, the ejaculates of normal and subfertile bulls were cryopreserved using semen extender in which there was IGF-1 supplementation at 0 (control), 50, 100, 150, 200, 250, 350 and 450 ng/mL of extender. Supplementation of IGF-1 at 250 ng/ml resulted in improved sperm motility, longevity and membrane intactness as compared to control after cryopreservation of semen from normal buffalo bulls, but not sub-fertile bulls. In summary, serum IGF-1 concentration was correlated with sperm mass motility and concentration in buffalo bulls and supplementation of IGF-1 protected sperm during the cryopreservation process but effects were not due to direct antioxidant activity.