The limited deformability and ATP depletion was considered by some authors to be the factor limiting the life span of old red blood cells (RBC) in circulation. Others believed that sialic acid on the RBC surface determines their life span. We compared the life span of 51Cr labelled, neuraminidase treated rabbit RBCs with ATP depleted by incubation at 37 degree C rabbit RBCs. Osmotic fragility, agglutinability, glucose-6-phosphate dehydrogenase (G6PD) and hexokinase activity and ATP levels of these cells were determined. Desyalated RBCs were removed from the circulation within 24 hours. ATP levels, G6d and hexokinase activity and osmotic fragility were normal in these cells. The agglutination by poly(L-lysine) was affected by the loss of surface charge on these cells. Half the ATP depleted RBCs were out of the circulation within three days. Reconstitution of ATP by reincubation with adenosine, elevated the ATP levels to about 80% of their original level, but survival of these cells did not improve. Analysis of sialic acid showed tha 50% of it was removed during the incubation for ATP depletion. The low ATP level and loss of sialic acid fromt he RBC membrane appeared to be conincidental rather than dependent on each other. The latter appears to be a primary factor in red cell survival.