Western blotting of IgG with extracts of normal human RBCs.

IgG purified from plasma of patients with WAIHA and healthy blood donors was immunoblotted at 200 μg/mL with a soluble extract of pooled RBCs of healthy donors, as described in Materials and Methods. Lanes 1 to 10: IgG of patients with WAIHA; lanes 15 to 24: IgG of healthy blood donors; lanes 12 and 13: normal reference IgG (IVIg); lanes 11 and 14: secondary anti-Fcγ antibody alone.