An effective methodology to isolate and characterize the Golgi complex of Tritrichomonas foetus is described in this work. Using sucrose density gradient centrifugation, two highly enriched Golgi fractions (GF1 and GF2) were obtained. Enzymatic assays of GF1 and GF2 showed a strong enrichment in galactosyltransferase activity (20- and 7-fold, respectively), with minimal contamination with other organelles. The GF fraction was further subfractionated by alkaline treatment, which resulted in the production of Golgi content and membrane subfractions. Electron microscopic observations of intact cells or Golgi fractions fixed in solutions containing glutaraldehyde and tannic acid, as well as of deep-etched replicas of isolated fractions, revealed the presence of discrete bridges only between closely apposed cisternae.