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To assess this, membrane was exposed to 5 mL of crude lysate from a standard OleD Loki E. coli heterologous overproduction culture for 24 h, subsequently washed with Tris-HCl buffer three times, and the purity of the captured protein upon release via treatment with 500 mM imidazole in Tris-HCl buffer solution was determined via SDS-PAGE, which revealed captured protein with a purity of ≥99%.
The buffer solution was stored at 4 °C before use.
No sheath flow of buffer solution is needed.
Degradation of sensor stored in buffer solution was evaluated.
The buffer solution was phosphate buffer (6 mM, pH 7.4).
A commonly used buffer solution is PBS (phosphate buffered saline solution).
The buffer solution is then collected or re-circulated.
At selected time intervals, buffer solution was taken for UV spectrometer and replaced with fresh buffer solution.
2.5 µl of 50 µM peptides solution were added close to the electrodes (2 ml of buffer) and images were captured with a CCD camera (Cool SNAP HQ) controlled with Metamorph software (Roper Scientific).
2.5 µl of 50 µM Penetratin solution were added close to the electrodes containing the GUV (2 ml of buffer) and images were captured with a CCD camera (Cool SNAP HQ) controlled with Metamorph software (Roper Scientific).
Both lipid and buffer solutions were fully degassed before loading into the appropriate cell.
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