Add time:08/26/2019         Source:sciencedirect.com

A simple and rapid method for determining the activity of dolichol monophosphate (cas 12698-55-4) phosphatase and dolichol diphosphate phosphatase is reported. This procedure is based on the separation of the substrates, [3H]dolichol monophosphate and [3H]dolichol diphosphate, from their reaction products, [3H]dolichol and [3H]dolichol monophosphate, by thin-layer chromatography on silica gel plastic plates. The chromatography chamber is of a nonconventional type, as it has a thin slot from which the last millimeter of the plate emerges. This special chamber allows the solvent to evaporate as it reaches the top of the plate. As a consequence, the mobile compounds tend to concentrate at the edge of the plate. The dolichol monophosphate phosphatase and dolichol diphosphate phosphatase substrates, which are more negatively charged than the corresponding reaction products, remain at the origin whereas the products migrate to the edge of the plate when appropriate solvents are used. The measurement of the radioactivity present at both the origin and the edge of the plate allows rapid estimation of the enzymatic activities.

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