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For imaging, 80 μl of fresh buffer solution was placed on the sample.
A reference cuvette filled with buffer solution was placed in the reference beam.
The homogenized buffer solution was placed on ice for 1 hour and then centrifuged at 4°C for 13,000 rpm for another 20 minutes, causing the supernatant solution to separate.
The homogenized buffer solution was placed on ice for 1 hour, centrifuged at 4°C for 13,000 rpm for another 20 minutes, and the supernatant solution was separated.
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All experiments were conducted in overflow mode at 20°C with proteins and ligands equilibrated in buffer A. 1.8 ml protein solution was placed in the temperature-controlled sample cell and titrated with the ligand loaded in the 300 μl mixing syringe.
To study the pH-stability of CauloGA, the enzyme solution was placed in various buffers (pH 3.5-11) at 4°C for 60 min, and the remaining activity was measured.
The solutions were placed in 6 8 kDa MWCO dialysis tubing and dialyzed against 2 L of 10 mM HEPES at pH 7.5 and 35 °C with daily buffer changes.
An equivalent volume (50 μl) of buffer solution was added to the blank or control in place of the extract.
The buffer solution was phosphate buffer (6 mM, pH 7.4).
A commonly used buffer solution is PBS (phosphate buffered saline solution).
The solution was mixed with equal volume of 66% sucrose in MBS buffer, the mixture was placed at the bottom of an ultracentrifuge tube, and a discontinuous gradient was formed by overlaying with the 30% sucrose and 5% sucrose solutions.
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