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For the histochemical demonstration of alkaline phosphatase, various methods such as azo dye method and cobalt calcium method have been employed. It has been difficult to check the precise localization of the enzyme on the tissue sections, because these reactions have been usually accompained with adsorption or diffusion of the reagents and the reaction products on tissues.We found a modified method by adding the high concentration of sodium chloride solution into the reagents mixture so that the precise localization could be obtained with minor diffusion and adsorption of the reagents and the products on the tissues.The mechanism of protection against adsorption and diffusion of the reagents and the products seemed to be mainly due to the salting-out effect by the sodium chloride. But it was not decided yet whether this phenomenon was truly due to the protection against adsorption and diffusion of the reagents and the products or was due to the positional variation of the disturbance of the enzyme activity by sodium chloride in tissues or cells. We have therefore biochemically and histochemically tried to explain the mechanism of the salt reaction on the activity of alkaline phosphatase.

Effects of Sodium Chloride on the Histochemical Demonstration of Non-Specific Alkaline Phosphatase and its Investigation