HeLa lysates were oxidized by the addition of H2O2 (hydrogen peroxide) for 0, 0.5 or 3.5 hours. Afterwards, cell lysates from the control or H2O2-treated samples were treated +/- dinitriphenylhydrazine, then separated by PAGE and transferred to a nitrocellulose membrane. Dinitrophenylated proteins were immunodetected using the anti-DNP antibody provided in the kit. The lanes contain: (1) dinitrophenylated molecular weight standards; (2) DNP-modified HeLa control cells; (3) DNP-modified, 0.5-hour H2O2-treated HeLa cells; (4) DNP- modified, 3.5 hour-H2O2-treated HeLa cells; (5) unmodified HeLa control cells; (6) unmodified HeLa, 0.5-hour H2O2-treatedHeLa cells. Reprinted with permission from Intergen.
A number of different processes result in the production of free radicals, which can then go on to modify macromolecules. Examples include metal-catalyzed oxidation, radiation-mediated oxidation, and oxidation via ozone or nitrogen oxides. The oxidative modification of proteins by these and other mechanisms results in the conversion of side chain groups of certain amino acid residues ...
