Figure 2.

Specificity of polyclonal antibody raised to NAL₂HNE_f (1a). As detailed in Materials and Methods, competitive ELISA was performed by using immunopurified anti-NAL₂HNE_f as primary antibody and NAL₂HNE_f-linked CA (1.0 pmol NAL₂HNE_f/well) as coating antigen. Antibody binding was evaluated in the presence of NAL₂HNE_f (○) and adducts produced in the reactions of HNE with N_α-acetylhistidine (▴), N_α-acetylcysteine (▪), and NAL (•), at concentrations indicated on the abscissa. B/B₀ = [(OD/s) − (OD/s)_bkg]/[(OD/s)_{no competitor} − (OD/s)_bkg] where OD/s = change in absorbance at 405 nm per s in the linear range. Background (bkg) values were obtained by using CA exposed to EDC in the absence of NAL₂HNE_f (1a) as coating antigen (Materials and Methods).