Human erythrocytes contain an acylphosphatase isoenzyme, whose concentration during cell ageing was determined by an enzyme immunoassay. Erythrocytes were age-fractionated by isopycnic centrifugation in "Percoll" density gradients. Acylphosphatase concentration was found to rise with the increase of cell density. Maximum values were attained in the mature erythrocytes and there was only a slight decrease in the older cells. Acylphosphatase activity in human erythrocytes of different ages followed a similar pattern, a finding which was confirmed for rabbit red cells with different levels of reticulocytes. The most probable mechanism for the increase of acylphosphatase content and activity during red cell maturation appears to be a de novo synthesis of this enzyme during the reticulocyte stage and its storage, virtually unaffected, in the mature erythrocytes. The possible consequences of increased acylphosphatase levels on the metabolic modifications associated with erythrocyte ageing are discussed.