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2 protocols using copper sulfate pentahydrate

1

Antioxidant-rich Lipid-Hyaluronate Formulation

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Lipoid S75 (consisting of ~70% of soy phosphatidylcholine, 9% phosphatidylethanolamine and 3% lyso-phosphatidylcholine) was purchased from Lipoid GmbH (Ludwigshafen, Germany). Sodium hyaluronate with low molecular weight (200–400 kDa) and a polydispersity of 1.4 Mw/Mn, was purchased from DSM Nutritional Products AG Branch Pentapharm (Switzerland). Tween 80, glycerol, DPPH radical (2,2-diphenyl-1-picrylhydrazyl), ferric chloride hexahydrate, gallic, chlorogenic, and ascorbic acids, catechin, tetrazolium salt, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), quercetin, neocuproin, Folin–Ciocalteu’s phenol reagent, 2,4,6-tris(2-pyridyl)-S-triazine (TPTZ) and pyrocatechol violet were purchased from Sigma-Aldrich (Milan, Italy). Potassium hexacyanoferrate (III) was obtained from Merck (Darmstadt, Germany). Anhydrous sodium acetate, copper sulfate pentahydrate and copper chloride dihydrate were obtained from VWR Chemicals BDH® (Frankfurt, Germany). Cyanidin 3-O-glucoside (≥95%) and delphinidin 3-O-glucoside (≥95%) were purchased from Extrasynthese (Lyon, France). Glacial acetic acid and phosphoric acid (85%) were acquired from J.T. Baker (Mallinckrodt Baker Inc., Utrecht, The Netherlands). All reagents and plastics for cell culture were purchased from Life Technologies Europe (Monza, Italy).
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2

Surface-Modified Fujifilm AEM Membranes

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Fujifilm AEM type I membranes
(Fujifilm Manufacturing Europe BV, The Netherlands) were used as base
membranes for surface modification. The membranes were conditioned
at room temperature and stored at 4 °C in 0.1 M NaCl solutions
before modification. N,N-Dimethylformamide
(DMF, 99.99%), absolute ethanol, and 2-propanol (IPA, 99.5%) were
used as solvents and purchased from Sigma-Aldrich (The Netherlands).
Dopamine hydrochloride (DA), triethylamine (TEA, ≥99.5%), 2-bromoisobutyril
bromide (BiBB), tris(hydroxymethyl) aminomethane (Tris), and [2-(methacryloyloxy)ethyl]dimethyl-(3-sulfopropyl)ammonium
hydroxide (known as sulfobetaine methacrylate, SBMA), copper(II) chloride, l-ascorbic acid, and sodium dodecylbenzenesulfonate (SDBS) were
also purchased from Sigma-Aldrich. Tris(2-pyridylmethyl)amine (TPMA)
was purchased from TCI Chemicals (Belgium). Hydrogen peroxide in 30%
solution, copper sulfate pentahydrate, and potassium hexacyanoferrate
(both ferri- and ferrocyanide, ≥ 96%) were purchased from VWR
Chemicals (Belgium). NaCl (99.5%) was purchased from ESCO (The Netherlands).
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