A "neutral" hexosaminidase has been separated from other hexosaminidase forms (I and II) by DEAE-cellulose chromatography and characterized in embryonic (16-days old) and 1-day old chicken brains. 2. Its properties differ from those of the forms I and II. It has optimum activity at about pH 6.0 and can be eluted from DEAE-cellulose with 0.25 M KCl only. 3. It has no N-acetylgalactosaminidase activity and cannot be successfully detected after isoelectric focusing since it is very acidic and completely unstable below pH 5.0. 4. "Neutral" hexosaminidase is heat-stable at pH 6.0 and is inhibited by chloride. 5. These properties, very different from those of forms I and II, suggest that this "neutral" form of hexosaminidase would be very similar to known hexosaminidase C separated from other materials. 6. We have found no significant differences for the above-mentioned three forms in chick embryos (16-days old) in comparison with those from 1-day old chicken.