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After which, a solution of saturated Na2CO3 (5 mL) was added.
After which, the reaction mixture was treated with a solution of saturated NH4Cl (5 mL).
After reaction completed, the reaction was quenched with a solution of saturated aqueous ammonium chloride (2 mL).
A solution of saturated aqueous solution of NaHCO3 (2 mL) was added and diluted with water (5 mL).
After cooling to rt, a solution of saturated aqueous NaHCO3 was added carefully to adjust the pH to slightly basic, and the mixture was heated for 15 30 min under reflux.
A 100 ml stock solution of saturated sodium chloride (salt solution) was prepared with distilled water and 38 g of NaCl; a 100 ml stock solution of 5M sodium hydroxide (base solution) was prepared with distilled water and 20 g of NaOH; and a 100 ml stock solution of 5M hydrochloric acid (acid solution) was prepared by diluting concentrated hydrochloric acid with distilled water.
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This solution was added to a solution of PPD saturated with glucose surfactant, and the resulting solution was subsequently heated in air to a reaction temperature of 60 120 °C.
Crystals were transferred to a cryo-protectant solution of (88% saturated Li2SO4, 14 mM magnesium formate, 20 mM Tris HCl (pH 8.0), and 0.1% (v/v) βME) by gradually changing the drop solution in 20 min, followed by cooling in liquid nitrogen.
The procedure was as follows: 100 μl of a 10 μg/ml solution of 50% saturated (NH4) 2S04 precipitate McAb HP10 diluted in carbonate-bicarbonate buffer 9.6 pH (Sigma C3041) was added to each of the wells of a flat bottomed Immunlon® 4HBX ELISA plate and the plate was incubated overnight at 4°C.
Table 1 Computed chemical properties of saturated CaCl2:2H2O solutions at 50-150°C 50-150°Cand
After the completion of the reaction, the mixture was washed successively with 1 M HCl, saturated solution of sodium bicarbonate and saturated brine solution.
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