✦ LIBER ✦

A batch procedure for the assay of cyclic nucleotide phosphodiesterase

✍ Scribed by Clark Bublitz

Publisher: Elsevier Science
Year: 1978
Tongue: English
Weight: 296 KB
Volume: 88
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(78)90402-5

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✦ Synopsis

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 the anion exchanger, which removes more than 95% of the tritium in boiled samples of [8-"HIcAMP or [VH]cGMP. Linear time courses and enzyme vs activity relationships are described for lo-" and 10-r M CAMP and 10e4 M cGMP. The method is limited by interference by neutral salts and by the enzymatic conversion of adenosine into inosine.

Methods

[8J4C]Inosine, 30 mCi/mmol, and [8-3H]cGMP, 0.5 Ci/mmol, were obtained from SchwarziMann.

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