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4 protocols using potassium hydrogen phosphate trihydrate

1

Quantitative Detection of Chicken Allergen Gal d 3

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Chicken ovotransferrin (Allergen, Gal d 3, purified from chicken egg white) and a matched antibody pair (unconjugated rabbit anti-Gal d 3 polyclonal antibody (CAb) and peroxidase-conjugated rabbit anti-Gal d 3 polyclonal antibody (DAb-HRP)) were supplied by MyBioSource. Bovine serum albumin (BSA), 3,3′,5,5′-Tetramethylbenzidine (TMB) liquid substrate (TMB-H2O2 K-Blue reagent), potassium dihydrogenphosphate, potassium hydrogenphosphate trihydrate, and sodium chloride were acquired from Sigma-Aldrich.
Phosphate-buffered saline (PBS), a 0.01 M phosphate solution containing 137 mM NaCl, pH 7.4, was used for both the biosensor construction (B1) and the allergen extraction (extraction buffer) from food samples. The daily working solutions of the biomolecules (standard allergen or food sample and DAb-HRP) were prepared in B1-BSA 0.5% (m/V).
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2

Biomimetic Mineral Deposition in Gelatin

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Gelatin type B (bovine skin, 225 Bloom), 1-ethyl-(3-3-dimethylaminopropyl carbodiimide hydrochloride) (EDC) and all reagents used to prepare SBF solution: Calcium chloride (CaCl2), Magnesium chloride hexahydrate (MgCl2·6H2O), Sodium bicarbonate (NaHCO3), Potassium hydrogen-phosphate trihydrate (K2HPO4·3H2O), Sodium sulfate anhydrous (Na2SO4), Potassium Chloride (KCl), Sodium chloride (NaCl) were purchased from Sigma-Aldrich (Milano, Italy).
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3

Histamine Dihydrochloride Analysis Protocol

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Histamine dihydrochloride (99%), sodium 1-decanesulfonate, potassium monophosphate, potassium hydrogen phosphate trihydrate, acetonitrile, and perchloric acid (HClO4) were purchased from Sigma-Aldrich (Amsterdam, The Netherlands). All chemical reagents and solvents were of analytical grade. Ultrapure water was obtained from a MilliQ purification system (Merck, Darmstadt, Germany).
Phosphate buffer solution at pH 6.9 was prepared by weighing 1.7 g of potassium monophosphate, 2.85 g of potassium hydrogen phosphate trihydrate, and 0.49 g of sodium 1-decanesulfonate and dissolving these in 1 L of distilled water.
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4

Supersaturated Calcification Coating Solutions

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Different supersaturated calcification coating solutions with high calcium and phosphate ion concentrations were prepared, based on the literature review [51 (link),52 (link),53 (link),54 (link)]. For comparative purposes, conventional simulated body fluid (SBF) with an ion concentration close to that of human blood plasma was prepared [55 (link)]. Each solution was prepared according to the reference (Table 1). Briefly, chemicals required for each solution were added one by one and completely dissolved in 800 mL of purified water at room temperature. Next, the volume of each solution was adjusted to 1 L using a volumetric flask. All solutions were prepared shortly before studies. The chemicals sodium chloride (NaCl), potassium chloride (KCl), sodium hydrogen carbonate (NaHCO3), calcium chloride dihydrate (CaCl2·2H2O), magnesium chloride hexahydrate (MgCl2·6H2O), sodium sulfate (Na2SO4), sodium dihydrogen phosphate monohydrate (NaH2PO4·H2O), and potassium hydrogen phosphate trihydrate (K2HPO4·3H2O) were used as received (>99.0% purity, analytical grade, Sigma-Aldrich). The ion concentrations of each solution are presented in Table 1.
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