19F NMR spectra were recorded at a frequency of 564.9 MHz on a Bruker AVANCE III HD ASCEND 600 MHz spectrometer equipped with BB(F) Double Channel Probe. All spectra were referenced relative to an external standard trifluorotoluene (TFT, –63.72 ppm). In phosphate buffers, NMR studies were performed at 25 °C, and in intraoocyte buffer, lysate, egg extract, and live oocytes, spectra were recorded at 18 °C. The following spectral parameters were used:48 (link),69 (link)
19F excitation pulse, 11 μs; spectral width, 21.28 ppm; transmitter frequency offset, –121.14 ppm; acquisition time, 1.4 s in phosphate buffer, 80 ms in intraoocyte buffer and lysate, 50 ms in egg extract and live oocytes; relaxation delay, 1.5 s in phosphate buffer, 1 s in intraoocyte buffer, lysate, egg extract, and live oocytes; number of scans 28 000 or 39 000. Using these parameters, 19F NMR spectra were obtained in 8–10 h. Each spectrum was processed with an exponential window function using lb = 10 Hz in case of studies in buffer, lysate, and egg extract and lb = 50 Hz in case of in-cell study.69 (link)
19F excitation pulse, 11 μs; spectral width, 21.28 ppm; transmitter frequency offset, –121.14 ppm; acquisition time, 1.4 s in phosphate buffer, 80 ms in intraoocyte buffer and lysate, 50 ms in egg extract and live oocytes; relaxation delay, 1.5 s in phosphate buffer, 1 s in intraoocyte buffer, lysate, egg extract, and live oocytes; number of scans 28 000 or 39 000. Using these parameters, 19F NMR spectra were obtained in 8–10 h. Each spectrum was processed with an exponential window function using lb = 10 Hz in case of studies in buffer, lysate, and egg extract and lb = 50 Hz in case of in-cell study.69 (link)
