Assay of dolichol monophosphate and dolichol diphosphate phosphatases: Thin-layer chromatography in a nonequilibrated system

A simple and rapid method for determining the activity of dolichol monophosphate 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]dolic...

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Autores principales: Boscoboinik, D.O., Belocopitow, E.
Formato: JOUR
Materias:
dolichol phosphate
phosphatase
dolichol diphosphate
enzyme assay
methodology
thin layer chromatography
Animal
Chromatography, Thin Layer
Kinetics
Microsomes, Liver
Phosphoric Monoester Hydrolases
Pyrophosphatases
Radioisotope Dilution Technique
Rats
Support, Non-U.S. Gov't
Tritium
Acceso en línea:http://hdl.handle.net/20.500.12110/paper_00032697_v114_n1_p42_Boscoboinik
Aporte de:
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
id todo:paper_00032697_v114_n1_p42_Boscoboinik
record_format dspace
spelling todo:paper_00032697_v114_n1_p42_Boscoboinik2023-10-03T13:55:46Z Assay of dolichol monophosphate and dolichol diphosphate phosphatases: Thin-layer chromatography in a nonequilibrated system Boscoboinik, D.O. Belocopitow, E. dolichol phosphate phosphatase dolichol diphosphate enzyme assay methodology thin layer chromatography Animal Chromatography, Thin Layer Kinetics Microsomes, Liver Phosphoric Monoester Hydrolases Pyrophosphatases Radioisotope Dilution Technique Rats Support, Non-U.S. Gov't Tritium A simple and rapid method for determining the activity of dolichol monophosphate 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. © 1981. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00032697_v114_n1_p42_Boscoboinik
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic dolichol phosphate
phosphatase
dolichol diphosphate
enzyme assay
methodology
thin layer chromatography
Animal
Chromatography, Thin Layer
Kinetics
Microsomes, Liver
Phosphoric Monoester Hydrolases
Pyrophosphatases
Radioisotope Dilution Technique
Rats
Support, Non-U.S. Gov't
Tritium
spellingShingle dolichol phosphate
phosphatase
dolichol diphosphate
enzyme assay
methodology
thin layer chromatography
Animal
Chromatography, Thin Layer
Kinetics
Microsomes, Liver
Phosphoric Monoester Hydrolases
Pyrophosphatases
Radioisotope Dilution Technique
Rats
Support, Non-U.S. Gov't
Tritium
Boscoboinik, D.O.
Belocopitow, E.
Assay of dolichol monophosphate and dolichol diphosphate phosphatases: Thin-layer chromatography in a nonequilibrated system
topic_facet dolichol phosphate
phosphatase
dolichol diphosphate
enzyme assay
methodology
thin layer chromatography
Animal
Chromatography, Thin Layer
Kinetics
Microsomes, Liver
Phosphoric Monoester Hydrolases
Pyrophosphatases
Radioisotope Dilution Technique
Rats
Support, Non-U.S. Gov't
Tritium
description A simple and rapid method for determining the activity of dolichol monophosphate 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. © 1981.
format JOUR
author Boscoboinik, D.O.
Belocopitow, E.
author_facet Boscoboinik, D.O.
Belocopitow, E.
author_sort Boscoboinik, D.O.
title Assay of dolichol monophosphate and dolichol diphosphate phosphatases: Thin-layer chromatography in a nonequilibrated system
title_short Assay of dolichol monophosphate and dolichol diphosphate phosphatases: Thin-layer chromatography in a nonequilibrated system
title_full Assay of dolichol monophosphate and dolichol diphosphate phosphatases: Thin-layer chromatography in a nonequilibrated system
title_fullStr Assay of dolichol monophosphate and dolichol diphosphate phosphatases: Thin-layer chromatography in a nonequilibrated system
title_full_unstemmed Assay of dolichol monophosphate and dolichol diphosphate phosphatases: Thin-layer chromatography in a nonequilibrated system
title_sort assay of dolichol monophosphate and dolichol diphosphate phosphatases: thin-layer chromatography in a nonequilibrated system
url http://hdl.handle.net/20.500.12110/paper_00032697_v114_n1_p42_Boscoboinik
work_keys_str_mv AT boscoboinikdo assayofdolicholmonophosphateanddolicholdiphosphatephosphatasesthinlayerchromatographyinanonequilibratedsystem
AT belocopitowe assayofdolicholmonophosphateanddolicholdiphosphatephosphatasesthinlayerchromatographyinanonequilibratedsystem
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