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Both buffer systems were prepared using newly developed precursor solutions containing a simple carboxylic acid as solvent.
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Systems were prepared in 10 × 10 × 10 Å buffered cubic box with a TIP3P solvent model.
"Systems are prepared," he said.
As a control, a second sample containing buffer and the Fd/NADP+ reductase system was prepared with no rTHB1 present ("–THB1") and was treated in the same manner as the sample containing rTHB1 ("+THB1").
Three independent samples of 50 nM CaMKII and 33 nM γP-ATP in Buffer System I were prepared and incubated on arrays, alongside a single negative control assay lacking kinase.
A second set of three independent samples of 50 nM CaMKII and 33 nM γP-ATP in the presence of 5 mM CaCl2 and 600 nM calmodulin, in Buffer System I was prepared and incubated on arrays, alongside a single negative control assay lacking kinase.
Buffer solutions were prepared using high purity water from a Millipore System (EASYpure RF, Barnstead, Germany).
All buffer solutions were prepared using ultrapure water (>18 MΩ cm) from a Millipore Milli-Q water purification system.
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).
Fresh buffer and standards were prepared daily.
The assay buffers were prepared in KCl-HCl buffers (pH 1.1 2.2) and sodium citrate acid buffers (pH 2.2 8.0).
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