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2 protocols using raw264.7 cell culture media

1

Antimicrobial and Anti-inflammatory Assays

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Brain Heart Infusion (BHI) broth and (BHIA) Mueller–Hinton (MH) broth for antimicrobial susceptibility testing were purchased from BD (Becton, Dickinson and company, Franklin Lakes, NJ, USA). RAW264.7 cell culture media and Dulbecco’s modified Eagle’s medium (DMEM) were purchased from Gibco (Grand Island, NY, USA). The MTT solution for cytotoxicity assay was purchased from Bio Basic (Amherst, NY, USA). Lipopolysaccharide (LPS), N-(1-naphthyl)-ethylenediamine, sulfanilamide, and phosphoric acid for anti-inflammatory activity were obtained from Merck (Billerica, MA, USA) and Sigma-Aldrich (St. Louis, MO, USA). 2,20-azinobis-(3-ethylbenzothiazolin-6 sulfonic acid) (ABTS), potassium persulfate, 6-hydroxy-2,5,7,8-tetramethyl-chroman-2-carboxylic acid (Trolox), and gallic acid monohydrate were purchased from Sigma-Aldrich (St. Louis, MO, USA). Folin–Ciocalteu reagent, TPTZ (2,4,6-tri(2-pyridyl)-s-triazine) and quercetin dehydrate were obtained from Merck (Billerica, MA, USA). Ethanol, methanol, sodium carbonate, aluminum chloride, potassium acetate, and formic acid for total phenolic and flavonoid detection, and HPLC assay were obtained from RCl Labscan (Bangkok, Thailand). All reagents and chemicals used were analytical grade.
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2

Antioxidant and Anti-inflammatory Assays

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Folin-Ciocalteu reagent, TPTZ (2,4,6-tri(2-pyridyl)-s-triazine), and quercetin dehydrate were purchased from Merck (Billerica, MA, USA). 2,2-diphenyl-1-picrylhydrazil (DPPH), 2,2′-azinobis-(3-ethylbenzothiazolin-6 sulfonic acid) (ABTS), 6-hydroxy-2,5,7,8-tetramethyl-chroman-2-carboxylic acid (Trolox), gallic acid monohydrate, resveratrol, oxyresveratrol, and lipopolysaccharide (LPS) from Escherichia coli O111:B4 were obtained from Sigma-Aldrich (St. Louis, MO, USA). RAW 264.7 cell culture media and Dulbecco’s Modified Eagle’s Medium (DMEM) were purchased from Gibco (Grand Island, NY, USA). 3-(4,5-dimethylthizaol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) was obtained from Bio Basic (Toronto, ON, Canada). Primary antibodies, anti-beta-actin monoclonal antibody, anti-iNOS/NOS II polyclonal antibody, and anti-COX-2 polyclonal antibody, as well as the peroxidase-conjugated secondary antibody were purchased from EMD Millipore corporation (Temecula, CA, USA).
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