This study was conducted to investigate the effects of protease supplementation on the behaviour, slaughter performance, meat quality, and immune organ indices of broilers. A total of 240 one-day-old Arbor Acres broiler chickens were randomly allocated to four treatments, with six replicates per treatment and 10 broilers per replicate. Chickens were fed corn-soybean meal basal diets supplemented with 0, 4, 8, and 12 IU/g of protease.Compared with 0 IU/g of protease supplementation, 12 IU/g of protease increased the average and minimum time of feeding and drinking behaviour (P < 0.05); increased the average, maximum, and minimum time of preening behaviour (P < 0.05); and decreased feeding, walking, and stretching frequency (P < 0.05). pH 24h of breast and thigh muscles had improved (P < 0.05). The shear force of the breast muscle and the cooking loss of the thigh muscle had decreased (P < 0.05). Compared with 0 IU/g of protease, 4 IU/g of protease increased the average and minimum time of lying behaviour (P < 0.05) and decreased stretching, scratching, and aggressive frequency (P < 0.05). The shear force of the thigh muscle and spleen index had enhanced (P < 0.05). Protease can be used as an additive in a broiler diet for production.