Fig. 2

The method development for reduced from β2GPI production and its comparison with native β2GPI. a Purified β2GPI could not reduced by cysteamine or GSH alone. b Purified β2GPI could reduced by thioredoxin-1 (TRX-1). c Reduced β2GPI blocked by GSH under reduced and non-reduced SDS-PAGE conditions. Purified β2GPI of the S-200 step (lane 2) was reduced by thioredoxin-1 (TRX-1) and then cysteine with cysteamine (lanes 3 and 4 with final concentrations of 40 and 80 mM, respectively, at 37 °C for 2 h) and GSH (lanes 5 and 6 with final concentrations of 40 and 80 mM, respectively, at 37 °C for 2 h) blockage (a). Purified β2GPI of S-200 step (lane 2) was reduced by thioredoxin-1 (TRX-1) and then cysteine with cysteamine (lanes 3 and 4 with final concentration of 40 and 80 mM, respectively, at 37 °C overnight) and GSH (lanes 5 and 6 with final concentration of 40 and 80 mM, respectively, at 37 °C for 2 h) blockage (b). The final comparison of commercial standard β2GPI (lane 2) with purified native (lane 3) and GSH blocked (lane 4) under reduced and non-reduced SDS-PAGE conditions. 5 μg protein were loaded into each lane