Localization of alkaline phosphatase separated electrophoreticahly in starch and acrylamide gels was achieyed by a modification of the Gomori method for this enzyme.Calcium phosphate l)recil)itates formed by enzymatic activity were converted to lead phosphate by immersion of gels in 0.08 M tris-maleate buffer, pH 7.0, containing 3 mM lead nitrate.After washing, treatment with ammonium sulfide produced visible Precil)itates of lead sulfide in the region occupied by the enzyme.Under the conditions used, this method showed a sensitivity of approximately 3 m .o1lI of phosphate.This compared fa 'orably with the azo coupling method tested and was approximately 20 times as sensitive as a method employing sodium alizarine sulfonate for the visualization of calcium phosphate ljrecipitates.
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Allen et al. (1963) studied this question.