With the evaluation of free radical scavenging activity, a novel antioxidant peptide (BHP) was isolated, purified and characterized from bovine hair. By using gel filtration chromatography (GFC) and high performance size exclusion chromatography (HPSEC), BHP was isolated and purified from bovine hair hydrolysates. Then, with the analysis of matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), BHP was identified to be a novel peptide with a major sequence of Cys-Glu-Arg-Pro-Thr-Cys-Cys-Glu-His-Ser (CERPTCCEHS) and a molecular weight of 1325.4 Da, which was mainly composed by 13 kinds of amino acids. Meanwhile, BHP exhibited the remarkable antioxidant activity to scavenge free radicals (ABTS and hydroxyl radicals) and inhibit the erythrocyte hemolysis of rat blood and lipid peroxidation of rat liver effectively. Furthermore, BHP was observed to possess the significant antioxidant ability to protect DNA and PC12 cells against the oxidative injury induced by hydrogen peroxide. All results of present study suggested that the antioxidant peptide from bovine hair could be used as a potential antioxidant agent to protect human health applied in pharmaceutical and medical industries. (C) 2015 Published by Elsevier Ltd.