2.5. NADPH-Dependent Peroxidase Activity Measurement
This protocol is extracted from research article:
Function and Regulation of Chloroplast Peroxiredoxin IIE
Antioxidants (Basel), Jan 21, 2021; DOI: 10.3390/antiox10020152

The reduction of peroxides by PRX-IIE was monitored with the GRX system as reductant. The activity was measured using a Cary 300 Bio UV/VIS spectrometer (Varian, Middelburg, The Netherlands) following NADPH oxidation at 340 nm. The assay was performed at 25 °C in quartz cuvettes with 2 µM PRX-IIE, 0.5 units glutathione reductase (GR), 200 µM NADPH, 1 mM EDTA, 1 mM GSH, varying amounts of GRX-S12 and peroxides (H2O2, t-BOOH, CuOOH) in 40 mM K-Pi, pH 7.2.

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