Three groups of isolated rat livers were perfused at 35 C with Krebs-Ringer bicarbonate buffer containing commercial bovine serum albumin (BSA) which had been purified by gel filtration on a column of Sephacryl S-200 and used within 12 hr of purification, or BSA which had been purified by gel filtration and stored at -70 C until used. The ability of livers to produce bile, retain potassium, and to maintain a constant level of glucose in the perfusate was greatly improved in the presence of purified albumin which had not been frozen. Such livers also showed the highest rates of urea synthesis, but the rate of release of aspartate aminotransferase (GOT) from cells and the bile salt content of the bile produced were similar to those found with unpurified BSA. Livers perfused with purified albumin which had been stored in the frozen state were slightly inferior to those perfused with nonfrozen albumin in their ability to produce bile and urea, to retain potassium and GOT within cells, and to maintain a constant concentration of glucose in perfusates. The concentration of bile salts in the bile produced by this group was also lower than that found with the other two groups. Overall, isolated rat livers benefited from perfusion with purified albumin, although freeze storage of this material rendered it slightly inferior to the nonfrozen material in its ability to support the liver.