Catalase from bovine liver

Gelatin (Type B; ca. 225 g bloom, bovine skin) was purchased from Sigma–Aldrich (St. Louis, MO, USA). Sodium hyaluronate (HA, MW ~1000 kDa) was purchased from Kewpie (Tokyo, Japan). Horseradish peroxidase (HRP, 140 U/mg), N-hydroxysuccinimide (NHS), N,N-dimethylformamide (DMF), hydrogen peroxide (H₂O₂) aqueous solution (31 w/w%), and catalase from bovine liver were purchased from Wako Pure Chemical Industries (Osaka, Japan). Water-soluble carbodiimide hydrochloride (WSCD·HCl) was obtained from Peptide Institute (Osaka, Japan). List of abbreviations used in this article is provided in Table 1.

Reagents were purchased from the following sources: 5,5-dimethyl-1-pyrroline N-oxide (DMPO) from Labotec (Tokyo, Japan); catalase (from bovine liver) from Wako Pure Chemical Industries (Osaka, Japan); 4-hydroxy-2,2,6,6-tetramethylpiperidine N-oxyl (TEMPOL) from Sigma Aldrich (St. Louis, MO), (+)-catechin from Tokyo Chemical Industry (Tokyo, Japan). GSE (Leucoselect^®) was obtained from Indena (Milan, Italy). According to the manufacture, Leucoselect^® is a grape seed extract with a well-defined chemical composition such as catechin-monomers and -oligomers, which was completely elucidated by instrumental analyses such as high performance LC-MS. This was also confirmed in a previous study in which Leucoselect^® was subjected to a LC-MS analysis showing that the calculated concentrations of (+)-catechin and (–)-epicatechin were 12.1% (wt/wt) and 6.6% (wt/wt), respectively [9 (link)]. All other reagents used were of analytical grade.

7,8-Dihdroxyflavone, 7-hydroxyflavone, 4′-hydroxyflavone, flavone, quercetin, naringenin, and 4-aminoantipyrine were purchased from Tokyo Kasei Kogyo Co., Ltd. (Tokyo, Japan). (+)-Catechin, pantethine, superoxide dismutase (from bovine liver), hemoglobin (from humans), and carboxypeptidase B were purchased from Sigma (Missouri, USA). 4′,5-Dihydroxyflavone was purchased from Alfa Aesar (Massachusetts, USA). CoA and peroxidase (from horseradish) were purchased from Oriental Yeast (Tokyo, Japan). Catalase (from bovine liver) was purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Resource Q, HiPrep DEAE FF 16/10, and a low molecular weight standard kit were obtained from GE Healthcare (Buckinghamshire, UK). Bio-Scale mini cartridges CHT Type I 40 µm media (5 ml) were obtained from Bio-Rad (California, USA). The “Protease inhibitor cocktail for use with mammalian cell and tissue extracts” was purchased from Nacalai Tesque (Kyoto, Japan). N-Ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline was purchased from Dojindo (Kumamoto, Japan). All other chemicals used were from commercial sources and of analytical grade. Porcine liver and blood were collected from the same individual.