An indirect, competitive enzyme-linked immunosorbent assay for the quantitation of carbamoyl-phosphate synthetase I (ammonia) in rat liver has been developed. Homogenization of the liver in 1% sodium deoxycholate is used for complete solubilization of the enzyme. The detergent does not interfere with the method if diluted to a concentration of 0.01% or lower. The assay is applied to determine the amount of enzyme in control rats and in rats fed "cafeteria" or high-protein diets. Changes in the amount of carbamoyl-phosphate synthetase I (ammonia) paralleled changes in enzymatic activity.