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2 protocols using isorhamnetin

1

Neurochemical Signaling Pathway Analysis

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Methanol, hexane, chloroform, ethyl acetate, butanol, lipopolysaccharide (LPS) from Escherichia coli serotype 026:B6 (Lot# 021M4072V), (-)-nicotine, methyllycaconitine citrate salt hydrate (MLA) from Delphinium brownii seeds, mecamylamine hydrochloride and 7-hydroxy-3H-phenoxazin-3-one-10-oxide sodium salt (resazurin) were purchased from Sigma-Aldrich (St Louis, MO, USA). Astragalin, baicalein, catechin, daidzin, daidzein, delphinidin, genistein, isoquercitrin, isorhamnetin, malvidin, petunidin-3-glycoside, quercetagetin, quercetin, rhamnetin, sakuranetin, spiraeoside and tamarixetin (Fig. 1) were purchased from Chromadex (Irvine, CA, USA). [3H]-MLA (∼60Ci/mmol), [3H]-cytisine (∼16Ci/mmol) and [3H]-epibatidine (∼30Ci/mmol) were purchased from American Radiolabeled Chemicals, Inc. (St. Louis, MO, USA). Antibiotics (10000U/mL penicillin and 10000ug/mL streptomycin), 2.5% trypsin (10×), fetal bovine serum (FBS), Dulbeccos' Modified Eagle Medium (DMEM), DMEM:Nutrient mixture F-12 (DMEM/F-12) and Hanks Balanced Salt Solution (HBSS) were purchased from Life Technologies Corporation (Grand Island, NY, USA). Griess reagent system was purchased from Promega Corporation (Madison, WI, USA). TNF-alpha ELISA Ready-SET-Go!® was purchased from eBioscience Inc. (San Diego, CA, USA).
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2

Analytical LC-MS Phenolic Standards

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Water, methanol, acetonitrile, 2-propanol and formic acid were of Optima® LC/MS grade (Fisher Scientific, Fair Lawn, NJ, USA). Glacial acetic acid provided by Merck (Darmstadt, Germany) was of Suprapur® quality. Sodium fluoride (Fluka Chemie, Buchs, Switzerland) and ascorbic and formic acid (Panreac, Barcelona, Spain) were of ACS grade.
The phenolic standards (+)-catechin, (-)-epicatechin, procyanidin B1, procyanidin B2, ferulic acid, quercetin-3-O-galactoside, isorhamnetin, isorhamnetin-3-O-glucoside and isorhamnetin-3-O-rutinoside were supplied by Extrasynthèse (Genay, France); procyanidin C1, quercetin-3-O-glucoside, quercetin-3-O-rhamnoside and quercetin-3-Oarabinofuranoside, by Chromadex (Santa Ana, CA, USA); caffeic acid, 5 ´-O-caffeoylquinic acid (5-CQA), p-coumaric acid, phloretin, phloretin-2′-O-glucoside and quercetin, by Sigma-Aldrich Chemie (Steinheim, Germany); phloretin-2′-O-xylosylglucoside, by Polyphenols Biotech (Bordeaux, France); and quercetin-3-O-rutinoside, by PhytoLab (Vestenbergsgreuth, Germany).
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