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Clinical Chemistry 29: 751-761, 1983;
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Clinical Chemistry, Vol 29, 751-761, Copyright © 1983 by American Association for Clinical Chemistry

A reference method for measurement of alkaline phosphatase activity in human serum

NW Tietz, CA Burtis, P Duncan, K Ervin, CJ Petitclerc, AD Rinker, D Shuey and ER Zygowicz

We present an official AACC reference method for the measurement of alkaline phosphatase, the culmination of optimization experiments conducted by a group of independent laboratories. The details of this method and evaluation of factors affecting the measurement are described. A metal ion buffer has been incorporated that maintains optimal and constant concentrations of zinc(II) and magnesium(II) ions. Final reaction conditions are: pH (30 degrees C), 10.40 +/- 0.05; 2- amino-2-methyl-1-propanol buffer, 0.35 mol/L; 4-nitrophenyl phosphate, 16.0 mmol/L; magnesium acetate, 2.0 mmol/L; zinc sulfate, 1.0 mmol/L; and N-(2-hydroxyethyl)ethylenediaminetriacetic acid, 2.0 mmol/L.


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