Nitric oxide (NO) is a product of the enzymatic conversion of arginine to citrulline by nitric oxide synthase. NO reacts rapidly with superoxide to form peroxynitrite. At physiological pH and in the presence of transition metals, peroxynitrite undergoes heterolytic cleavage to form hydroxyl anion and nitronium ion, the latter of which nitrates protein tyrosine residues. Thus, the presence of nitrotyrosine on proteins can be used as a marker for peroxynitrite formation in vivo. The presence of nitrotyrosine has been detected in various inflammatory processes including atherosclerotic placques.
NO tyrosine antibody
Western blot - Nitro tyrosine antibody (ab42789)Image courtesy of Gwen Lagoda by Abreview.
All lanes: Whole cell lysate prepared from human penile tissue loaded at 30µg. Gel is 4-15% mini gel, performed under reduced, denaturing conditions. ab42789 added at a 1/1000 dilution. Secondary used was a donkey anti-rabbit IgG conjugated to HRP, used at a 1/7000 dilution.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Nitro tyrosine antibody (ab42789)Image from Sánchez A et al., PLoS One. 2012;7(4):e36027. Epub 2012 Apr 23. Fig 6.; doi:10.1371/journal.pone.0036027; April 23, 2012, PLoS ONE 7(4): e36027.
Immunohistochemical analysis of prediabetic obese rat penile tissue, staining Nitro tyrosine with ab72789.
Samples were blocked with 10% normal goat serum, permeabilized with 0.3% Triton X-100 before being incubated with primary antibody (1/100) for 48 hours at 4°C. A biotinylated goat anti-rabbit IgG (1/400) was used as the secondary antibody and staining was detected using DAB.
Timpani CA et al. Attempting to Compensate for Reduced Neuronal Nitric Oxide Synthase Protein with Nitrate Supplementation Cannot Overcome Metabolic Dysfunction but Rather Has Detrimental Effects in Dystrophin-Deficient mdx Muscle. Neurotherapeutics14:429-446 (2017).
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Fernandes CR et al. High-Fat and Fat-Enriched Diets Impair the Benefits of Moderate Physical Training in the Aorta and the Heart in Rats. Front Nutr4:21 (2017).
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