Minor Hemoglobins in Erythrocytes

FREE-BOTJNDARY ELECTROPHORESI8 has been used for the isolation, from fresh extracts of normal human red blood cells, of a hemoglobin, which at pH 8.8 was more negatively charged than the chief component (1) and which, on spectroscopic examination, has been found indistinguishable from the bulk of the oxyhemoglobin as regards the visible region and the Soret band (2). The proportion of this hemoglobin to the total hemoglobin seemed to be 2.5 per cent, after a correction has been applied for the boundary anomalies. Two hemoglobins, differing from the bulk hemoglobin by their electrophoretic mobilities, have recently been isolated by zone electrophoresis in a starch slab (3). One of them, migrating faster than the chief component, was probably identical with the hemoglobin previously described (1). The other hemoglobin was more positively charged than the chief component and migrated more slowly. Its concentration ranged from 1.8 to 3.5 per cent of the total hemoglobin. The demonstration of the “fast” hemoglobin and the quantitative determination of the “slow” hemoglobin of human red cells by electrophoresis on paper has been accomplished and is the subject of the present paper.