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The tissues were frozen in liquid nitrogen and stored at -80°C until preparation for solution hybridisation analyses.
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Accordingly, a suggested standardized sample preparation for CAP solutions in an EtOH-based solvent system is detailed below.
The methods for the preparation of solutions and for all enzyme assays in 30 mM TEA buffer (pH 7.5) at I = 0.1 (NaCl) and 25 °C were described in a recent publication.
SS (Standard Solution) - containing deionized water, 1 mM KCl, 1 mM CaCl2 and 50 mM mannitol (pH 7 – TRIS/MES) - as a control for the injection method and the base for preparation of solutions of active substances.
All water used for the various steps of the hybridization and for preparation of solutions was filtered (0.2 μM) MilliQ dH20.
Double-distilled deionized water was used for preparation of solutions.
UO2Ac2·2H2O salt (Chemapol, Czech Republic) was used for preparation of solutions.
Tb NO3 3·6H2O were prepared by dissolving the corresponding oxides in diluted nitric acid, and nanopure water was used for preparation of solutions.
For preparation of solutions containing ferric ions, Fe(NO3 3 was dissolved in deionized water or artificial saliva (80 mM).
Doubly distilled deionized water (DDW) was used for the preparation of solutions, wherever required.
All the chemicals used were of analytical reagent grade obtained from Merck (Mumbai, India), and deionized water is used for the preparation of solutions.
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