Figure 3.

Binding of anti-NAL₂HNE_f to HNE-treated Glu-6-PDH. As detailed in Materials and Methods, the binding of immunopurified anti-NAL₂HNE_f to Glu-6-PDH-linked Lys-HNE fluorophore (60 fmol/well) was evaluated by ELISA. In addition, antibody binding was evaluated in the presence of NAL₂HNE_f (1a) (○), fluorophores produced in the reactions of HNE with γ-aminobutyric acid (1c) (□) and ethylamine (1e) (▵), and nonderivitized NAL (•), γ-aminobutyric acid (▪), and ethylamine (▴). B/B₀ = [(OD/s) − (OD/s)_bkg]/[(OD/s)_{no competitor} − (OD/s)_bkg] where OD/s = change in absorbance at 405 nm/s in the linear range. Background (bkg) values were obtained by using Glu-6-PDH treated with HNE for 0 min (Materials and Methods).