Siliaflash f60

Reagents were purchased from Sigma-Aldrich, VWR, Acros Organics, or Tokyo Chemical Industry (TCI) and used as received. Spectroscopic grade, inhibitor-free THF was deoxygenated by sparging with argon followed by passage through a Pure Process Technologies solvent purification system to remove water. Deuterated solvents were purchased from Cambridge Isotope Laboratories and used as received. Silica gel (SiliaFlash F60, Silicycle, 230–400 mesh) was used for column chromatography. ¹H, ¹³C{¹H}, and ¹⁹F NMR spectra were recorded on a Bruker 500 or 600 MHz instrument (as indicated). Chemical shifts are reported in parts per million relative to residual protic solvent resonances. Mass spectrometric measurements were performed by the University of Illinois, Urbana–Champaign MS facility. All air-free manipulations were performed under an inert atmosphere using standard Schlenk techniques or an Innovative Atmospheres N₂-filled glovebox. HTCM-NO₂, HTCM-F, and HTCM-Me were prepared as described in the literature, with spectral data matching those reported previously.^{30 (link)}

Reagents were purchased from Sigma Aldrich, Tokyo Chemical Industry (TCI), Fisher Scientific, and VWR and used directly as received. Carbonic anhydrase (CA) from bovine erythrocytes was purchased from Sigma-Aldrich (C2624). Silica gel (SiliaFlash F60, Silicycle, 230−400 mesh) was used for column chromatography. Deuterated solvents were purchased from Cambridge Isotope Laboratories (Tewksbury, Massachusetts, USA). ¹H, ¹⁹F, and ¹³C{¹H} NMR spectra were recorded on Bruker 500 MHz NMR instrument at the indicated frequencies. Chemical shifts are reported in ppm relative to residual protic solvent resonances. Mass spectrometric measurements were performed by the University of Illinois, Urbana Champaign MS facility, or on a Xevo Waters ESI LC/MS instrument. Methylene blue absorbance was monitored by an UV−vis spectrometer (Cary 100, Agilent Technologies, Santa Clara, California, USA) in PBS buffer. SF7-AM,^{71 (link)}ATB-346,^{72 (link)} and iminothioether 2²⁵ were synthesized by following the literature report. HeLa cells were purchased from ATCC (Manassas, Virginia, USA). Cell imaging experiments were performed on a Leica DMi8 fluorescence microscope, equipped with an Andor Zyla 4.2+ sCMOS detector.

Reagents were purchased from Sigma-Aldrich, Tokyo Chemical Industry (TCI), Fisher Scientific, and VWR and used directly as received. Carbonic anhydrase (CA) from bovine erythrocytes was purchased from Sigma-Aldrich (C2624). Silica gel (SiliaFlash F60, Silicycle, 230–400 mesh) was used for column chromatography. Deuterated solvents were purchased from Cambridge Isotope Laboratories (Tewksbury, Massachusetts, USA). ¹H, ¹⁹F and ¹³C{¹H} NMR spectra were recorded on Varian INOVA 500 MHz, Bruker 500 MHz, or Bruker 600 MHz NMR instruments at the indicated frequencies. Chemical shifts are reported in ppm relative to residual protic solvent resonances. H₂S Detection was monitored by using an H₂S electrode (ISO-H₂S-2, World Precision Instruments, Inc. Sarasota, Florida, USA) or UV-Vis spectrometer (Cary 60, Agilent Technologies, Santa Clara, California, USA) by following methylene blue assay in PBS buffer. Stability test was performed on HPLC (1260 Infinity II, Agilent Technologies, Santa Clara, California, USA). OA-PeroxyTCM-1, OA-PeroxyTCM-2, OA-PeroxyTCM-3, and OA-TCM-1 were synthesized by following our previous study.⁶⁹

Reagents were purchased from Sigma-Aldrich, Tokyo Chemical Industry, or VWR and used directly as received. Deuterated solvents were purchased from Cambridge Isotope Laboratories and used as received. ¹H, ¹³C{¹H}, and ¹⁹F NMR spectra were recorded on a Bruker 500 MHz instrument. Chemical shifts were reported relative to residual protic solvent resonances. MS data was collected on a Xevo G2-XS QTof (Waters) instrument. Silica gel (SiliaFlash F60, Silicycle, 230–500 mesh) was used for column chromatography. All air-free manipulations were performed under an inert atmosphere using standard Schlenk technique.