Antibodies to nitrotyrosine were prepared in goats or rabbits by injecting a nitrotyrosine–protein conjugate. The antibodies were purified by using a protein that had been nitrated with tetranitromethane. These antibodies were used to isolate nitrotyrosine-containing peptides from nitrated lysozyme. The nitrotyrosine-containing peptides were thus purified (55% yield) in one step and the positions of nitration in lysozyme were found to be at tyrosine-20 and tyrosine-23. This method is of general applicability for the determination of the position of nitrotyrosine in proteins.

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