A Rapid and Economical Method of Preparing Radioiodinated Cyclic Nucleotide Derivatives for use in Radioimmunoassays
| Autor: | S. W. Kubalak, M. G. Currie, W. F. Oehlenschlager |
|---|---|
| Rok vydání: | 1990 |
| Předmět: |
Pharmacology
chemistry.chemical_classification Chromatography Immunology Radioimmunoassay Guanosine Reversed-phase chromatography Amino acid Iodine Radioisotopes chemistry.chemical_compound Cyclic nucleotide chemistry Isotope Labeling Chloramine-T Cyclic AMP Tyrosine Bioassay Nucleotide sense organs Cyclic GMP Sodium acetate |
| Zdroj: | Journal of Immunoassay. 11:109-118 |
| ISSN: | 0197-1522 |
| DOI: | 10.1080/01971529008053262 |
| Popis: | 2'-O-succinyladenosine 3':5'-cyclic monophosphate tyrosyl methyl ester (ScAMP-TME) and 2'-O-succinylguanosine 3':5'-cyclic monophosphate tyrosyl methyl ester (ScGMP-TME) were radioiodinated using chloramine T and Na125I. The resulting radiolabeled cyclic nucleotide derivatives, ScAMP-125I-TME and ScGMP-125I-TME, were subsequently purified by reverse-phase chromatography on Sep-Pak C18 cartridges (Waters Associates, Milford, MA) and tested as tracers in sensitive radioimmunoassays for cAMP and cGMP, respectively. Purified ScAMP-125I-TME and ScGMP-125I-TME functioned in the respective radioimmunoassays for up to 12 weeks when suspended in a 1:1 (v:v) mixture of n-propanol and 20 mM sodium acetate, pH 6.0. Thus, this purification method enables rapid and economical preparation of tracers for cyclic nucleotide radioimmunoassays. Furthermore, our findings suggest that reverse-phase chromatography may be applicable to the purification of other small polar molecules to which tyrosyl groups have been added for the purpose of radioiodination. |
| Databáze: | OpenAIRE |
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