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Pre-treatment of HGF with trisulfides and nitration Recombinant mouse HGF (carrier protein-free) was pre-treated with glutathione trisulfide (GSSSG) and lipoic acid trisulfide (LASSS) at mole ratios (HGF : trisulfide) of 1:0 (with solvent alone), 1:400, 1:1000, and 1:4000 at pH 7.4, 25C just prior to the peroxynitrite protocol (optimized at a molar ratio 1:2000 relative to HGF, pH 7.4, 37, for 30 min), which was previously demonstrated to induce maximal nitration of Y198 and Y250 located in the -chain under physiological conditions as described previously 32
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Intramuscular concentrations of anserine in these non-primate species range from 2 mM for opossum to 21 mM for marlin
Over-dilution (too much water) increases oxidative exposure surface area
