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All buffer solutions were treated with biotechnology-grade Chelex 100 (143-2832, Bio-Rad).
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All buffers and protein solutions were treated with Chelex resin to ensure the complete removal of contaminating calcium.
The PCR solutions were treated with DpnI.
The resultant mixed sample solution was treated with fixed concentration of nitric acid followed by pH adjustment between 3.0 and 6.0 using buffer solution.
Three different buffer solutions were prepared: pH 4 citrate buffer 0.1 M, pH 6 citrate buffer 0.1 M and PBS 1x (pH 7.4).
Both lipid and buffer solutions were fully degassed before loading into the appropriate cell.
Octanol and buffer solutions were presaturated.
To survey our cholesterol-derived oxysterols for protein reactivity, buffer solutions of HSA were treated with each of the alkynyl oxysterol electrophiles shown in Figure 1.
Thus, buffer solutions of HSA were treated with a-seco A at varying concentrations (0 100 μM), and biotinylated proteins were visualized using the streptavidin fluorophore, with the integrated intensities showing that the amount of protein lipidation increases with increasing concentration of alkynyl lipid electrophile.
Each incubation vial contained 12 islets in 1.0 ml KRB buffer solution and was treated with 95%O2 5%% CO2 to obtain constant pH and oxygenation.
Paraffin sections were deparaffinized through xylene for three changes of five minutes each, followed by graded alcohol immersions to water and phosphate buffered saline solutions, the sections were treated with the retrieval solutions (DAKO, Dakocytomation Inc.,Copinteria,CA,USA) for 20 minutes at 95°C.
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