USE OF AMP (2-AMINO-2-METHYL-1-PROPANOL) BUFFER FOR HISTOCHEMICAL DEMONSTRATION OF ALKALINE PHOSPHATASE ACTIVITY

抄録

AMP (2-amino-2-methyl-1-propanol) buffer was tried for the histochemical detection of alkaline phosphatase (ALPase) activity by the lead citrate method. The incubation medium tested was as follows: 170-350mM AMP, 20mM sodium β-glycerophosphate, 3.9mM magnesium sulfate, 2.0mM lead citrate and 8% sucrose; pH was adjusted to 9.2. By using this medium, ALPase activity in the liver was found in the sinusoidal and lateral surface of hepatocytes as well as in the bile canalicular surface, which was the only positive site by the original lead citrate method using Tris buffer. Moreover, the reaction product (lead phosphate) appeared finer than when using the Tris method. By quantitating ALPase activity biochemically, higher activity was found with AMP than with Tris. Apparently, this new incubation medium with AMP is suitable for detecting weak ALPase activity.