Haptoglobin Genotype Is a Determinant of Hemoglobin Adducts and Vitamin E Content in HDL

Figure 3

Hydroxyurea addition to HDL, Hb, and H₂O₂ inhibits Hb cross-link formation. HDL (obtained from the plasma of healthy volunteers), Hb, and H₂O₂ (1 : 10 ratio) were incubated with different concentrations of hydroxyurea for 1 hour. The samples were loaded on SDS-PAGE gel, transferred to a PVDF membrane, and stained with anti-Hb antibody. The sample order is (1) HDL, (2) HDL + Hb + H₂O₂, (3) HDL + Hb, (4) Hb, (5) Hb + H₂O₂, (6) HDL + Hb + H₂O₂ + hydroxyurea (400 nM), (7) HDL + Hb + H₂O₂ + hydroxyurea (200 nM), (8) HDL + Hb + H₂O₂ + hydroxyurea (100 nM), and (9) HDL + Hb + H₂O₂ + hydroxyurea (10 nM).