Isoelectric Focusing of Alkaline Phosphatase Isoenzymes in Polyacrylamide Gels: Use of Triton X-100 and Improved Staining Technic

Abstract

The examination of alkaline phosphatase isoenzymes by means of isoelectric focusing in polyacrylamide gel rods using the apparatus and focusing method of Righetti and Drysdale is discussed. A simultaneous coupling procedure using α-naphthyl phosphate and fast blue salt R in 2-amino-2-methyl-1,3-propanediol buffer, pH 9.68, containing MgCl₂ and ZnSO₄ proved sensitive for developing the enzyme bands. Also discussed are the effects seen with the incorporation of Triton X-100 into the gel and sample mixtures. Enzyme, which remained at the top of the gel without using this detergent, entered the gel easily with the addition of Triton X-100 into the application solution. Incorporation of Triton into the gel matrix resulted in some enzyme band patterns that showed distinct differences from gels containing no Triton.