Fig. 2.

Effect of reaction pH on peroxynitrite-induced modifcation of perlecan. Surface absorbed perlecan (10 nM) was treated with peroxynitrite (10 μM) at 22 °C for 20 min in 0.1 M phosphate buffer, pH 6 – 7.5 in the absence (black bars) and presence (light grey bars) of NaHCO₃ (25 mM). The perlecan was then probed by ELISA using mAbs against HS epitopes; (A) mAb 10E4 and protein core epitopes; (B) perlecan core (mAb CSI-076), (C) perlecan domain III (mAb 7B5) and (D) perlecan domain V (mAb CSI-074). Data are expressed as % normal ELISA signal and are means ± SEM of values obtained from triplicate wells with n ≥ 3. Data were analyzed by 2-way ANOVA, with statistical significance assumed at p < 0.05, a different to perlecan control, b different to dONOO, * effect of NaHCO₃.