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roGFP2 fluorescence was recorded for 18 h at 25 C in a Synergy H1 multi-detection microplate reader (Biotek, Bad Friedrichshall, Germany) at the excitation wavelengths 405 and 488 nm and emission wavelength at 525 nm with the following settings: 20 nm bandwidth, gain 75 %, intervals: 1 min 41 sec, optics: bottom

Generally, it performs best in aqueous systems within a pH range of 5.0 to 7.0