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5 protocols using aluminium trichloride

1

Extraction and Analysis of Phytochemicals

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Organic solvents used for extraction were purchased from Sigma Aldrich (Castle Hill, NSW, Australia). Other chemicals and standards of analytical grade or higher were also sourced from Sigma Aldrich, such as 2,2′-diphenyl-1-picrylhydrazyl (DPPH), 2′-azino-di-(3-ethylbenzthiazoline sulfonic acid) (ABTS), 3-(2-Pyridyl)-5,4,4-dimethoxybenzaldehyde,4-triazine-p,5,6-diphenyl-1,6-hydroxy-2,6-Tris(2-pyridyl)-s-triazine (TPTZ), 7,8-tetramethylchroman-2-carboxylic acid (Trolox), aluminium trichloride, anhydrous sodium carbonate, catechin hydrate, disodium ethylenediaminetetraacetic acid (EDTA-Na2), ferrous chloride, Folin–Ciocalteu reagent, gallic acid monohydrate, phloroglucinol, potassium persulfate, p’-disulfonic acid monosodium salt hydrate (Ferrozine), sodium acetate, trisodium phosphate, and quercetin. HPLC grade standards including phloroglucinol, gallic acid, chlorogenic acid, syringic acid, synaptic acid, catechin, epicatechin, and epigallocatechin were purchased from Sigma Aldrich (Castle Hill, NSW, Australia). Vanillin was obtained from Chem-Supply Pty Let., Adelaide, SA, Australia. The Milli-Q water used was obtained from Millipore Milli-Q Gradient Water Purification System (Darmstadt, Germany).
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2

Antioxidant Activity Determination

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The polyphenolic standards (gallic acid, hydrated quercetin-3-rutinoside), H2O2, ABTS peroxidase and trolox were provided by Sigma-Aldrich (Berlin, Germany). The reagents (Folin-Ciocalteu, aluminium trichloride, acetate of sodium) were provided by Sigma-Aldrich (Berlin, Germany), and the other solvents were obtained from Panreac (Barcelona, Spain), and Lab-Scan (Warsaw, Poland).
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3

Phytochemical Analysis and Antioxidant Activity

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All analytical grades of dissolvent, reagents and pure phenolic compounds such as Methanol, Folin–Ciocalteu Reagent, sodium carbonate (Na2CO3), Aluminium Trichloride (AlCl3), Hydrochloric Acid (HCl), Gallic Acid (C7H6O5), Catechin (C15H14O6), Quercetin (C15H10O7), Vanillin (C8H8O3), Sodium Phosphate Monobasic (NaH2PO4), Sodium Dibasic Phosphate (Na2HPO4), Sulfuric Acid (H2SO4), Potassium persulfate (K2S2O8), Iron chloride (II) (FeCl2), Iron Trichloride (FeCl3), Ammonium acetate, Sodium hydroxide (NaOH), Potassium Hydroxide (KOH), 1,1-Diphenyl-2-picryl- hydrazyl (DPPH), β-carotene, Linoleic Acid, Tween 40, Trichlo-roacetic acid (TCA), Potassium ferricyanide (K3Fe[CN]6), Ammonium molybdate (NH4) 2MoO4, Acetylcholinesterase (AChE) type VI-S from electric eel < 1000 U/mg solid, Acetylcholine iodide (ACI), 5,50-dithiobis (2-nitrobenzoic acid) (DTNB), Carragenean, Indometacin, Omepazole, Galanthamine, Butylated hydroxytoluene BHT (C15H24O), Butylate hydroxyanisole BHA (C11H16O2) and Ascorbic Acid (C6H8O6) were purchased from Sigma-Aldrich and Fluka.
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4

Cytotoxicity and Oxidative Stress Assays

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Media and all cell culture related reagents were purchased from Biosera (Kansas City, MO, USA). Sulforhodamine-B (SRB) and trichloro-acetic acid (TCA) were purchased from Fluorochem (Glossop, UK), whereas Trisma Base, Aluminium trichloride and sodium acetate were purchased from Sigma-Aldrich (Merk) (St. Louis, MO, USA). TBARS and Protein Carbonyl Colorimetric Assay Kits were purchased from Cambridge Bioscience Ltd. (Cambridge, UK). The Folin–Ciocalteu Phenolic content assay kit was purchased from Bioquochem (Parque Tecnológico de Asturias, Llanera, Spain), whereas the Caspases - 3, - 8 and - 9 multiplex activity assay kit was purchased from Abcam (Cambridge, UK). All solvents were of UHPLC optima grade or better.
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5

Alzheimer's Disease Biomarker Assay

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Aluminium trichloride, urethane, hydrogen peroxide (Sigma-Aldrich, St. Louis, MO, USA), maltolate (Sigma-Aldrich, St. Louis, MO, USA), anti-phospho-CREB (Ser133) (Cell Signalling, USA), anti-phospho-tau (Ser396), anti-PKA (Affinity, USA), anti-PRKACB (Merck, USA), anti-tau (Sercivebio, CN), anti-GAPDH mouse, goat anti-rabbit immunoglobulin (IgG), goat anti-mouse IgG, miRNA extraction kit, high sensitivity chemiluminescence liquid ECL, protein quantitative kit, tissue protein extraction kit, lane marker loading buffer (Reducing, 5x), EasyQuick RT MasterMix, Acry (Beijing ComWin Biotech Co., Ltd. CN), all-in-one miRNA qRT-PCR detection kit, primer for miR-200a-3p, primer for U6 (FulenGen, CN), DYC Z-24DN electrophoresis tank DYCP-40C semi-dry membrane transfer instrument (Liuyi instrument factory, Beijing), Bio-Rad chemiluminescence imaging system (Bio-Rad, USA), and ethyl carbamate (Sigma-Aldrich, St. Louis, MO, USA) were used in this study.
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