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3 protocols using sodium hydroxide solution

1

HPLC Analysis of Anticancer Drugs

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Standards of ABE, RIB, and PAL were purchased from Toronto Research Chemicals (Toronto, ON, Canada). Benzimidazole (BEN) was purchased from Sigma-Aldrich (Burlington, MA, USA). Tablets of ABE (Verzenios) and RIB (Kisqali), as well as capsules of PAL (Ibrance), which are clinically available, were used. Running buffers were prepared from 50 mM sodium phosphate solution with pH = 2.5 and 1 M sodium hydroxide solution, both purchased from Agilent Technologies (Santa Clara, CA, USA). LC-grade acetonitrile and methanol were purchased from VWR International (Radnor, PA, USA). Formic acid was purchased from Kemika (Zagreb, Croatia). Ultra-pure water used for the preparation of all solutions and running buffers was obtained using a Milli-Q system.
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2

Optimization of Phytochemical Extraction

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Hydroxytyrosol, tyrosol, vanillin, vanillic acid, ferulic acid, p-coumaric acid, cinnamic acid, luteolin, and apigenin were used as reference standards for the optimization of the extraction procedure and purchased from Sigma–Aldrich (Schnelldorf, Germany). methanol (HPLC-grade quality) was used for the extraction (Baker, Avantor Performance Materials, Arnhem, Netherlands). For HPLC–Qtof–MS analysis formic acid (Honeywell Fluka, Fisher Scientific, Schwerte, Germany), methanol (Fisher Scientific, Schwerte, Germany), isopropanol (99.9%, Honeywell, Riedel-de-Häen, Fisher Scientific, Schwerte, Germany), 1 M sodium hydroxide solution (Agilent Technologies, Santa Clara, CA, USA), and water (Merck, Darmstadt, Germany) were used (HPLC-grade quality).
Petroleum ether (40–60 °C analytical grade > 98%), heptane, tocopherols standards, sodium methylate, sodium hydrogen sulphate (monohydrate, extra pure), and tert-butyl methyl ether (HPLC grade) were purchased from Merck (Darmstadt, Germany). Tocopherol and tocotrienol standard compounds were purchased from CalBiochem (Darmstadt, Germany)
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3

Protein-DNA Conjugation Protocol

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Thrombin from human plasma and apo-transferrin were purchased from Sigma-Aldrich (USA). BSA and fibrinogen from human plasma were purchased from Wako Pure Chemicals Industries (Japan). All ssDNAs, except for the random ssDNA library with its primer set, were synthesized by Sigma-Aldrich (USA). 2-Morpholinoethanesulfonic acid (MES; Wako, Japan), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimido hydrochloride (EDC; Wako, Japan), Dulbecco’s PBS(−) (Wako, Japan), 10% Tween 20 solution (Bio-Rad, USA), tris(hydroxymethyl)aminomethane (Tris; Wako, Japan), 1 M sodium chloride solution (Wako, Japan), 1 M magnesium chloride solution (Wako, Japan), 1 M sodium hydroxide solution (Agilent Technologies, USA), 0.5 M borate buffer at pH 8.5 ± 0.2 (Polysciences, USA), EDTA disodium salt dihydrate (Wako, Japan), and boric acid (Wako, Japan) were used as received. All solutions were prepared using ultrapure water from a Milli-Q water purification system (Merck Millipore, USA). For the preparation of gels, a 37.5:1 (40%, w/v) acrylamide/bis solution, 2.6% C (Serva Electrophoresis, Germany), ammonium persulphate (Bio-Rad, USA), and N,N,N’,N’-tetramethylethylene (TEMED; Bio-Rad, USA) were used. The loading buffer and a 25-bp DNA stepladder were purchased from Wako (Japan).
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