Three Golgi fractions, GF-1, GF-2 and GF-3, were isolated from the livers of rats pretreated with colchicine, which gave better yields of the fractions than ethanol treatment of rats. 2. Electron microscopic observation showed that GF-1 was composed mainly of secretory vesicles, GF-3 consisted predominantly of small tubules and flattened cisternae, and GF-2 was an intermediate fraction composed of secretory vesicles and cisternal elements. 3. Among these three fractions the highest activity of galactosyl transferase, marker enzyme of the Golgi complex, was found in GF-3 and the lowest activity was in GF-1, although a different distribution of the enzymes was observed in fractions obtained from ethanol-treated rat liver. 4. Enzymatic characterization of these fractions showed that no significant contamination with other subcellular components occurred in GF-1 and GF-2.