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. Author manuscript; available in PMC: 2012 Dec 17.
Published in final edited form as: Free Radic Biol Med. 2012 Jan 8;52(6):1075–1085. doi: 10.1016/j.freeradbiomed.2011.12.024

Fig. 9.

Fig. 9

Exposure of J774A.1 cells to HOSCN results in the formation of protein-derived sulfenic acid species. J774A.1 cells (1 × 106 cells ml−1) were pretreated with DAz-2 (5 mM in 1% v/v DMSO) or DMSO (1% v/v) for 30 min at 37 °C before the addition of PBS or HOSCN (50–100 µM) and further incubation for 15 min at 37 °C. The cells were then lysed, and Staudinger ligation was carried out with p-biotin (200 µM) and DTT (5 mM) for 2 h at 37 °C. The reaction was quenched and proteins were separated by 1D SDS–PAGE (4–12%). Proteins (50 µg per lane) were then transferred to PVDF membranes and biotinylated proteins detected by HRP–streptavidin. Asterisks denote endogenously biotinylated proteins.