Xanthine oxidase xod from bovine milk

AETS, TTAU, HTAU, and cysteamine hydrochloride were kindly gifted from Sogo Pharmaceutical Co., Ltd. (Tokyo, Japan). 5,5-Dimethyl-1-pyrroline-N-oxide (DMPO) was purchased from Labotec (Tokyo, Japan), xanthine oxidase (XOD) from bovine milk was from Sigma-Aldrich (St. Louis, MO), hypoxanthine (HX) and iron(II) sulfate heptahydrate were from FUJIFILM Wako Pure Chemical Co. (Osaka, Japan), heparin sodium salt solution was from Mochida Pharmaceutical Co., Ltd. (Tokyo, Japan), ethylenediamine-N,N,N’,N’-tetraacetic acid, disodium salt, dihydrate (EDTA) and diethylenetriamine-N,N,N’,N’’,N’’-pentaacetic acid (DTPA) were from Dojindo (Kumamoto, Japan). Pure water was freshly prepared with a Millipore Milli-Q Labo (Bedford, MA). All other reagents were of the highest purity commercially available.

MEM and antibiotics (streptomycin and penicillin) were from HyClone, USA. Fetal bovine serum (FBS) was from Gemini Bio-products, USA. 3-(4, 5-dimethylthiazol-2yl)-2, 5-diphenyltetrazolium bromide (MTT) and Xanthine oxidase (XOD, from bovine milk) were purchased from Sigma-Aldrich Chemicals Co. (St. Louis, MO, USA). 2,6-dihydroxyacetophenone (DHAP, 99%), 4-Hydroxyphenethyl alcohol (4-HPEA, 98%), and 4-Hydroxybenzaldehyde (HBA, 98%) were purchased from Macklin Biochemical Co., Ltd (Shanghai, China). Xanthine and allopurinol were purchased from Shanghai Macklin Biochemical Co., Ltd. Dimethyl sulfoxide (DMSO) was obtained from Xilong Scientific Co., Ltd (Guangdong, China). All reagents were analytical grade. The deionized water was obtained from the Milli-Q Reagent Water System of Milliore Co. (Milford, MA).