LOCALIZATION OF ALKALINE PHOSPHATASES IN GEL MATRICES FOLLOWING ELECTROPHORESIS

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Abstract
Localization of alkaline phos-phatase, separated electrophoretically in starch and acrylamide gels, was achieved by a modification of the Gomori method for this enzyme. Calcium phosphate precipitates, formed by enzymatic activity, were converted to lead phosphate by immersion of gels in 0.08 [image] tris-maleate buffer, pH 7.0, containing 3 m[image] lead nitrate. After washing, treatment with ammonium sulfide produced visible precipitates of lead sulfide in the region occupied by the enzyme. Under the conditions used, this method showed a sensitivity of approximately 3 m[mu][image] of phosphate. This compared favorably 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 precipitates.

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