A New Method for the Preparation of Band 3, the Main Integral Protein of the Human Erythrocyte Membrane

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Abstract
Band-3 protein from human erythrocyte membranes was isolated, without using detergents, by a 2-step procedure. The peripheral proteins were removed from the membrane by treatment with 10% acetic acid. The remaining lipoprotein complex was solubilized in approximately 92% (vol/vol) acetic acid and then separated into its components by preparative zonal electrophoresis in a gradient made up of acetic acid, water and sucrose. Band 3 was recovered from the gradient at a yield of 60-70% and a purity of about 95%. Approximately 25 mg of band 3 could be prepared in 1 run. The protein is soluble in aqueous solutions, even in the absence of organic solvents or detergents. In addition to band 3, the proteins stained by periodic and/Schiff''s reagent (the sialoglycoproteins) are also separated from the other proteins.