A Batch Elution Procedure for Assaying Adenylate Cyclase
โ Scribed by S.P.J. Brooks; M.J. Mancebo; C.P. Holden; K.B. Storey
- Publisher
- Elsevier Science
- Year
- 1993
- Tongue
- English
- Weight
- 286 KB
- Volume
- 210
- Category
- Article
- ISSN
- 0003-2697
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๐ SIMILAR VOLUMES
who provided us with some details of this salt procedure, unpublished at that time.
A procedure for the assay of cyclic nucleotide phosphodiesterase is described in which labeled cyclic nucleotide is separated from labeled nucleoside by the batchwise addition of ethanolic slurries of Dowex 2 fluoride. Under the conditions described there is no detectable adsorption of nucleoside by
548) has been modified by using [3H]ATP to produce [3H]cyclic AMP and monitoring the recovery with [14C]cyclic AMP. The modified assay remains sensitive and accurate, but avoids the inconvenience and expense of using 32p.
In the assay of adenylate cyclase using [&\*P]ATP as the substrate and alumina chromatography as the separating procedure for labeled nucleotides, blank levels are dependent on the quality of the labeled ATP and also on that of the alumina. In order to lower the blanks by eliminating the radioactive