Acrylamide-Gel Electrophoresis of Hemoglobins

Abstract

Abstract Details of a technic for analysis of hemoglobin mixtures using acrylamide-gel electrophoresis are presented. The method offers increased speed and reproducibility. Measurements on the unstained gel are made at 525 and at 420 m µ in a procedure affording increased precision of measurements for minor components. The gel is then stained with amidoblack to detect and measure nonhemoglobin components. The major component of normal human adult hemoglobin is separated into two components in this medium. The two components (A1, A3) appear in varying relative amounts depending on the source of the specimen. The normal minor component A2 also appears. After staining the pattern with a protein dye such as amidoblack 10B, additional components appear in the electrophoresis patterns of hemolysates from carefully washed red cells. These components have a constant migration ratio referred to hemoglobin A1 and appear to be nonheme-containing intraerythrocytic proteins.