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. 2013 Apr 8;14(4):7542–7570. doi: 10.3390/ijms14047542

Figure 3.

Figure 3

Effects of bovine aortic microsomes on the nitration of bovine serum albumin (BSA) and of Ellman’s reagent on the nitration of PGIS and other proteins by peroxynitrite. (A) Detection of 3-nitrotyrosine in BSA by Western blot analysis using a monoclonal anti-3-nitrotyrosine antibody. BSA (5 μM) was incubated with authentic peroxynitrite (PN, 0–250 μM) in the presence or absence of bovine aortic microsomes (1 mg/mL total protein); (B) HPLC analysis of free 3-nitrotyrosine content in pronase digests from samples used for Western blot; (C) Detection of 3-nitrotyrosine positive proteins in bovine aortic microsomes by Western blot analysis using a monoclonal anti-3-nitrotyrosine antibody. Bovine aortic microsomes (1 mg/mL total protein), which were either treated or not with the sulfhydryl oxidizing Ellman’s reagent (DTNB) were mixed with authentic peroxynitrite (PN, 0–100 μM). Data are representative of two independent experiments.