LOCALIZATION MECHANISMS IN ENZYME CYTO-CHEMISTRY STUDIED WITH ALKALINE PHOSPHATASE-LOADED ERYTHROCYTE-GHOSTS

Erythrocyte ghosts containing varying amounts of alkaline phosphatase were used to study the localization mechanisms of 3 metal salt and 1 azo method for this enzyme. For the azo method, the minimal amount of alkaline phosphatase that can be visualized within the ghosts proved only to be limited by the optical properties of the azo compound. In contrast, for the metal salt methods, a certain threshold activity had to be present in the ghosts in order to obtain correct localization of the final reaction product. The localization properties of both azo and metal salt methods conformed to the theories of cytochemical enzyme localization presented to date. By determining the rate constant of the capture reaction and the diffusion constant of the primary product, the localization properties of the azo method could be predicted. Some remaining discrepancies between theory and practice are discussed.