Figure 1.

Nitrotyrosine antibody specificity. Competition for nitrotyrosine (NT) antibody (Ab) binding to a NT containing 13mer peptide coated on a plate. ELISA competitors are free nitrotyrosine (diamond), NT containing 33mer peptide (circle), chlorotyrosine (square), and tyrosine (triangle). Competition for Ab binding was most effective with free NT followed closely by the NT containing 33mer peptide. This shows that the Ab will recognize free NT as well as NT containing proteins in biological fluids. Significant competition by tyrosine and modified tyrosine occurred at greater than two orders of magnitude higher concentrations of each, indicating very good NT Ab specificity.

Figure 2.

Western blot using OBR’s nitrotyrosine immunoblot positive control on nitrocellulose with the nitrotyrosine antibody at 1:1,000 dilution. Lane 1, 2.44 µg; Lane 2, 9.75 ng positive control (nitrated carbonic anhydrase).