Western blot analysis of membrane-associated 4.1R.

Schematic representation of erythroid domains and corresponding cDNA structure for the normal 4.1R and CO.1 missense C-terminal domain is shown. The asterisk indicates the premature termination codon in CO.1. Red cell membrane proteins were resolved with SDS-PAGE and transferred to a nitrocellulose membrane. Each sample corresponds to 20 μg of total membrane protein. The antibodies used in this set of Western blot studies are depicted beneath the structural domains, except that Ab4.1 epitopes are not indicated precisely because this polyclonal antibody was raised against the whole 80-kd isoform. The tested samples were as follows: heterozygous 4.1R Coimbra (family member B2: 1, 7, 10, and 13), homozygous 4.1R Coimbra (family member B1: 6, 9, and 12), heterozygous 4.1R Fier (family member A1: 5), heterozygous 4.1R Presles (4), and control membrane samples with normal 4.1R (2, 3, 8, and 11).