Fig. 1.

Chromatograms of human RBC lysate as analyzed by reverse phase chromatography and Western blot analysis. Reverse phase HPLC was performed using an YMC-pack PROTEIN-RP column (4.6 × 250 mm) as described in Section 2. The separated proteins were collected in 1.0 ml fractions and concentrated using a centrifugation concentrator system (Sakuma, Tokyo, Japan) for further analysis. After concentration, all fractions (3–50 min) were subjected to Western blot analysis with anti-Prx2 antibody after 12% SDS–PAGE and transfer to a PVDF membrane. RBC lysates were prepared after treatment with 500 μM H₂O₂ (B), t-BHP (C) or without peroxide (A). A typical result from the other three samples providing similar results is presented.