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3 protocols using methionine

1

Purification and Assay of Porcine Cu/ZnSOD

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Cu/ZnSOD from porcine erythrocytes was purchased from Biodee Biotechnology Co., Ltd. 2-Mercaptobenzimidazole (MBI), nitroblue tetrazolium (NBT), methionine, riboflavin and EDTA were obtained from Sinopharm Chemical Reagent Co., Ltd. A 0.2 molL−1 mixture of phosphate buffer (mixture of NaH2PO4·2H2O and Na2HPO4·12H2O, pH = 7.4) was used to control the pH. All other reagents were of analytical grade and purchased from standard reagent suppliers. Ultrapure water (18.25 MΩ) was used throughout the experiments.
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2

Antioxidant Activity Evaluation of Green Coffee

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Hydroxyhydroquinone (99%), L-cysteine, 2-furfurylthiol (98%), 1,2dichlorobenzene (99%) and ethanol were purchased from Sigma-Aldrich Co. Ltd (Shanghai, China). Disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium sulfite, iron (Ⅲ) chloride hexahydrate (FeCl3•6H2O), iron (Ⅱ) sulfate monohydrate (FeSO4•H2O), hydrogen peroxide (H2O2, 30%), egg albumin, ethylenediaminetetraacetic acid (EDTA, disodium salt), ethyl acetate, ascorbic acid, methionine and glutathione (99%) were purchased from Sinopharm Chemical Reagent Co. Ltd (Shanghai, China). Millipore water was made by Ultrapure Water Maker Type 1 (Weston, USA). Green Robusta coffee beans (sun drying, crop years 2016/2017) from Vietnam was purchased from Royal Coffee Co. Ltd (Shanghai, China).
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3

Comprehensive Antioxidant Assays Protocol

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RNA extraction kit was obtained from Beijing TransGen Co., Ltd., China, while qRT-PCR kit and reverse transcription kit were purchased from Beijing Takara Co., Ltd., China.
Chemical reagents (analytical grade), such as dibasic sodium phosphate (Na2HPO4), ferric chloride, potassium ferricyanide, ascorbic acid (AsA), hydrogen peroxide (H2O2), sodium diethyl dithiocarbamate trihydrate, sodium dihydrogen phosphate (NaH2PO4), polyvinyl pyrrolidone (PVP), 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxylamine hydrochloride, α-naphthyl amine, p-aminobenzenesulfonic acid, sodium nitrite, hydroxylamine, trichloroacetic acid, thibabituric acid, dithiothreitol, nitro blue tetrazolium, riboflavin, ethylenediaminetetraacetic acid disodium salt (EDTA-Na2), bovine serum albumin, catechin, gallic acid, methionine, dithio nitrobenzoic acid (DTNB), glutathione (GSH), methanol, were obtained from Shanghai Sinopharm Co. Ltd., China.
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