To investigate the anti-diabetic and antioxidant effects of Opuntia humifusa cladodes, O. humifusa cladodes powder was extracted with 75% ethanol and fractionated with various solvents. Among the extracts fractionated with various solvents, ${alpha}$-amylase and ${alpha}$-glucosidase inhibitory activities of O. humifusa cladodes were highest ($89.94{pm}1.15%$ and $29.01{pm}3.03%$, respectively) in the ethyl acetate fraction. Further, total phenolic and flavonoid contents of the ethyl acetate fraction were the highest ($196.02{pm}5.26$ and $114.00{pm}10.03{mu}g/mg$, respectively). DPPH and ABTS radical scavenging activities increased according to the concentration of O. humifusa cladodes extract, and those of the ethyl acetate fraction were the highest. Ferric reducing antioxidant powers of chloroform and the ethyl acetate fraction were higher than those of other fractions. Overall, the ethyl acetate fraction of O. humifusa cladodes showed the highest anti-diabetic and antioxidant effects. Results indicate that O. humifusa cladodes powder has potential as a useful ingredient with anti-diabetic and antioxidant effects.