𝔖 Bobbio Scriptorium
✦   LIBER   ✦

A method for assaying orotate phosphoribosyltransferase and measuring phosphoribosylpyrophosphate

✍ Scribed by D.J. Hupe; N.D. Behrens

Publisher
Elsevier Science
Year
1987
Tongue
English
Weight
485 KB
Volume
161
Category
Article
ISSN
0003-2697

No coin nor oath required. For personal study only.

✦ Synopsis

An orotate phosphoribosyltransferase, OPRTase, assay method which relies upon binding reactant ['Hlorotic acid and product ['H]orotidine-S-monophosphate to polyethyleneimineimpregnated-cellulose resin and collecting on a GFC glass fiber filter is presented. Elution with 2 X 5 ml of 0.1 M sodium chloride in 5 mM ammonium acetate removes all of the orotate and leaves all of the product orotidine monophosphate (OMP) bound so that it may be measured in a scintillation counter. It was found that the addition of 10 ELM barbituric acid riboside monophosphate to the reaction mixture prevented the conversion of OMP to UMP and products of UMP. The assay is suitable for measurement of OPRTase activity with purified enzyme or in crude homogenates. A modification of this scheme using commercially available yeast OPR-Tase and 10 pM of unlabeled OMP provides an assay for phosphoribosylpyrophosphate with a sensitivity such that 10 pmol of PRPP may be measured. o 1987 Academic press, IX.

πŸ“œ SIMILAR VOLUMES

A microdetermination method for assaying
A microdetermination method for assaying glycosaminoglycans and proteoglycans
✍ P.Mark Bartold; Roy C. Page πŸ“‚ Article πŸ“… 1985 πŸ› Elsevier Science 🌐 English βš– 422 KB

A simple and reliable method is described for the determination of glycosaminoglycans and proteoglycans in tissue extracts as well as during preparative and analytical procedures for these molecules. It is particularly useful because it requires much less starting material for the identification of

A Nonradioactive High-Performance Liquid
A Nonradioactive High-Performance Liquid Chromatographic Microassay for Uridine 5β€²-Monophosphate Synthase, Orotate Phosphoribosyltransferase, and Orotidine 5β€²-Monophosphate Decarboxylase
✍ Jerapan Krungkrai; Nuchanat Wutipraditkul; Phisit Prapunwattana; Sudaratana R. K πŸ“‚ Article πŸ“… 2001 πŸ› Elsevier Science 🌐 English βš– 76 KB

A novel nonradioactive, microassay method has been developed to determine simultaneously the two enzymatic activities of orotate phosphoribosyltransferase (OPRTase) and orotidine 5'-monophosphate decarboxylase (ODCase), either as a bifunctional protein (uridine 5'-monophosphate synthase, UMPS) or as

A method for measuring microquantities o
A method for measuring microquantities of DNA
✍ GiachΓ¨, Valentino ;Pirami, Laura ;Becciolini, Aido πŸ“‚ Article πŸ“… 1994 πŸ› John Wiley and Sons βš– 320 KB

Quantitative analysis o f DNA content represents a critical step when only very small amounts of nucleic acids are available. The DNA content of a small RNA-free sample can be measured in a simple and precise way using a two-dimensional approach. DNA samples are spotted on the surface o f an agarose