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Quantitative starch-gel zone electrophoresis of human hemoglobins



Quantitative starch-gel zone electrophoresis of human hemoglobins



Acta Pharmaceut Jugoslav 13(2): 69-76



In separating different types of human hemoglobin, many methods have been used, including alkaline denaturation and solubility techniques, chromatography, moving boundary electrophoresis, and zone electrophoresis on filter paper, agar, potato starch-grain and starch-gel. Electrophoresis is technically the simplest, very rapid and most suitable for preparative work among all methods available. Though different hemoglobins behave similary in an electric field on paper or starch-gel at pH 86, the latter one has a pronounced advantage by separating more clearly individual hemoglobins, making possible to differentiate the 2 unpigmented fractions X and X2. Till now hemoglobin A2 could be quantitatively easier determined in a starch-block. Using starch-gel electrophoresis, hemolized erythrocytes of normal persons, with pernicious anemia and j3-thalassemia were investigated. The starch-gel was cast as a very thin layer (3 to 4 mm) so that it was possible to prepare afterwards a plastic, translucent film for direct den-sitometry. In normal persons the hemolizates showed the following fractions: A3 + A1, A2 X and X2; in pernicious anemia and [beta]-thalassemia however a completely distinct fraction of fetal hemoglobin was found also.

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