A Quantitative Histochemical Study of Alkaline Phosphatase Activity in Isolated Rat Duodenal Epithelial Cells

摘要: A simple separation method enabling the quantification of alkaline phosphatase activity in unfixed, isolated, individual, duodenal epithelial cells has been presented. The intestinal brush border-bound demonstrated using naphthol AS-BI phosphate as a substrate and hexazotized New Fuchsin simultaneous coupling agent. amount final reaction product, measured cytophotometrically, increases linearly with incubation time (up to 10 min) concentration 0.4 mM). Maximum enzyme was obtained at pH 8.9. Variation revealed kinetic parameters for Km = 0.17 ± 0.015 Vmax 13.9 1.38. specificity confirmed by complete inhibition presence l-cysteine (10 mm) 80% L - phenylalanine (30 Comparison 8-μm cryostat sections (beginning tip proceeding cryptal part) along villus axis, individual successive separation, similar values percentage quotient derived from entire activities these two different methods. This suggests that presented procedure gives rise isolation respective corresponding areas villus. Finally, isolated can be used valuable tool quantitative analysis length