Deoxygenation of alligator red blood cells (RBCs) caused binding of two HCO3- equivalents per hemoglobin (Hb) tetramer at physiological pH. At lowered pH, some HCO3- binding also occurred to oxygenated Hb. The erythrocytic total CO2 content was large, and Hb-bound HCO3-, free HCO3-, and carbamate contributed about equally in deoxygenated cells. The nonbicarbonate buffer values of RBCs and Hb were high, and the Hb showed a significant fixed acid Haldane effect. Binding of HCO3- on deoxygenation occurred without a change in RBC intracellular pH, revealing equivalence between oxylabile HCO3- and H+ binding. Erythrocyte volume, plasma pH, and plasma HCO3- concentration also varied little with the degree of oxygenation. Diffusional water permeability was higher in oxygenated than deoxygenated RBCs. The RBCs have rapid band 3-mediated Cl- and HCO3- transport, which was not affected by degree of oxygenation, but net fluxes of Cl- and HCO3- via the anion exchanger are small during blood circulation at rest. Most of the CO2 taken up into the blood as it flows through tissue capillaries is carried within the erythrocytes as Hb-bound HCO3- until CO2 is excreted when blood flows through pulmonary capillaries.