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. Author manuscript; available in PMC: 2016 Feb 29.
Published in final edited form as: Methods Enzymol. 2014 Dec 26;552:185–210. doi: 10.1016/bs.mie.2014.10.007

Figure 5.

Figure 5

dPRX1/Jafrac1 behaves like a classic 2-cys peroxiredoxin. Drosophila PRX1/Jafrac1 was expressed, purified and H2O2-treated as in (Peskin et al., 2013) with the following modifications: 50 mM TCEP was used for the initial reduction step prior to desalting, 5 μM recombinant protein was used in a total volume of 50 μL for each concentration, and the incubation time with H2O2 prior to catalase (marked with an asterisk on the SDS-PAGE) addition and NEM treatment was 15 minutes. PRX identity and oxidation state was confirmed by mass spectrometry, antibody activity against the over-oxidized recombinant protein was confirmed by immunopreciptation/mass spectrometry. Marker used: MagicMark XP (Life Technologies).