Fig. 8.

Immunoblotting for HBF showing structural changes to human plasma FN treated with a MPO/H₂O₂/Cl⁻ system and increasing concentrations of SCN⁻. Purified human plasma FN (0.2 μM in 0.1 M phosphate buffer, pH 7.4) was either left untreated (MPO only control) or treated with MPO (0.02 or 0.1 μM), Cl⁻ (100 mM), H₂O₂ (160 μM) and increasing concentration of SCN⁻, and incubated for 2 h at 37 °C. Samples were electrophoresed on SDS-PAGE under A) non-reducing or B) reducing conditions, transferred onto PVDF membranes and probed with a mouse monoclonal anti-FN HBF antibody (A32; 1:5000), and conjugated with anti-mouse HRP secondary antibody (1:2000). Blots were developed with ECL-plus reagent. Black arrow = dimer/higher aggregates; white arrow = monomer band.