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Clinical Chemistry 18: 548-553, 1972;
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Clinical Chemistry, Vol 18, 548-553, Copyright © 1972 by the American Association for Clinical Chemistry

New Procedure for Purifying and Crystallizing Alkaline Phosphatase from Human Intestinal Mucosa

Sheshadri Narayanan 1 and Harold D. Appleton 1

1 Department of Clinical Chemistry, Metropolitan Hospital Center and Department of Biochemistry, New York Medical College, New York, N.Y. 10029.

A new procedure is described for extracting, purifying, and crystallizing alkaline phosphatase orthophosphoric monoester phosphohydrolase (EC 3.1.3.1) from human intestinal mucosa. Active enzyme, extracted with trichlorotrifluoroethane, is subsequently purified by chromatography on Sephadex G-200 and DEAE Sephadex. The acetone and ammonium sulfate precipitation steps that are part of the conventional butanol extraction and purification procedure are eliminated. Enzyme activity is concentrated at each stage of the purification procedure, yielding a preparation with a specific activity three times greater than that obtained with the conventional butanol procedure.


Key Words: trichlorotrifluoroethane extraction • Sephadex-gel permeation • anion-exchange chromatography • immunodiffusion • chromatography on DEAE Sephadex

Submitted on March 10, 1972
Accepted on April 7, 1972






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Copyright © 1972 by the American Association for Clinical Chemistry.