Skip to main content
. Author manuscript; available in PMC: 2014 Mar 1.
Published in final edited form as: Free Radic Biol Med. 2012 Nov 7;56:64–71. doi: 10.1016/j.freeradbiomed.2012.10.550

Figure 4.

Figure 4

ELISA for DMPO adduct detection on polynucleotides oxidized by the Cu2+ and H2O2 in the presence and absence of DMPO. Polynucleotides were: poly(dG).poly(dC), poly(dG-dC). poly(dG-dC), poly(dA).poly(dT), and poly(dA-dT).poly(dA-dT). Polynucleotides (100 µg mL−1), 20 µM Cu2+, 500 µM H2O2and 100 mM DMPO in PBS were incubated for 1 h at 37 °C. DNA (0.5 µg/well) was bound to an ELISA plate overnight in a Reacti-Bind DNA coating solution (Pierce). DMPO adducts were detected with a mouse monoclonal anti-DMPO antibody (panel a) or a chicken polyclonal anti-DMPO antibody (panel b). Results are means ± SD of triplicate measurements and representative of at least three independent experiments. One-way ANOVA with Dunnett’s Multiple Comparison Test showed the treatments that produced a significant increase over the untreated polynucleotides as indicated (* p <0.05).