Figure 2.

Preincubation experiment with sodium sulfite and horseradish peroxidase/H2O2. 10 mM Na2SO3 were preincubated (15 min, room temperature) with 50 mM DMPO. Then, 50 mM the other DMPO isotope, 1 mg/ml HRP and 10 μM H2O2 were added and the ESR spectrum was recorded immediately. A: Preincubation with 14N-incubation and addition of 15N-DMPO, enzyme and H2O2. B: Preincubation with 15N-DMPO; thereafter, addition of 14N-DMPO, HRP and H2O2. Both signals are almost identical and the differential spectrum (C; A minus B) shows no significant signal. The artifact formed according to the Forrester-Hepburn mechanism does not contribute to the ESR signal in a significant manner.