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2 protocols using cell culture medium

1

Antioxidant Compound Evaluation Protocol

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Naringenin, lycopene, β-carotene, hydroxytyrosol, 1,1′-dioctadecyl-3,3,3′,3′ tetra-methylindocarbocyanine perchlorate (Dil), p-nitro blue tetrazolium (NBT), phorbol myristate-acetate (PMA), superoxide dismutase (SOD), catalase, and fucoidan were obtained from Sigma Chemical Co. (St. Louis, MO, USA). Naringenin 7-O-β-d-glucuronide was obtained from Cayman Chem. Co. (Ann Arbor, MI, USA). Cell culture medium, foetal bovine serum and supplements were obtained from GE Healthcare Hyclone (Logan, UT, USA). All other reagents were of analytical grade. Assayed compounds were dissolved in dimethylsulfoxide (DMSO) and the final concentration of DMSO in cell cultures was lower than 0.1%.
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2

Exosome Proteomic Profiling by TMT

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Reagents and kits were purchased from commercial sources. The Total Exosome Isolation Reagent was purchased from Invitrogen. Iodoacetamide (IAA), dithiothreitol (DTT), urea and cell culture medium were from GE Healthcare (Little Chalfont, Bucks, UK). The BCA protein assay kit, TMT Mass Tagging Kits and foetal bovine serum (FBS) were from Thermo Fisher Scientific, and protease inhibitor cocktail was from Roche. Sequencing-grade endoproteinase Trypsin/Lys-C was from Promega. Transwell Permeable Support 8.0 μm Polyester (PET) Membrane 6.5 mm Insert and Matrigel Basement
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