A radioligand-binding assay for adenosine in tissue extracts |
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| Authors: | Ray A. Olsson Charles J. Davis Mary K. Gentry Robert B. Vomacka |
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| Affiliation: | 1. Department of Cardiorespiratory Diseases, Walter Reed Army Institute of Research, Washington, D. C. 20012 USA;2. Suncoast Heart Association Cardiovascular Research Laboratory, Department of Internal Medicine University of South Florida College of Medicine, Tampa, Florida 33620 USA |
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| Abstract: | A radioligand-binding assay employing adenine analog-binding protein from rabbit crythrocytes and [3H]adenosine of high specific activity measures less than 1 pmol of adenosine in neutral HClO4 extracts of tissue. Adenine and its nucleotides interfere with adenosine binding, so are removed by preliminary chromatography on PEI and phenyldihydroboryl cellulose columns. Free and bound adenosine are separated by filtration through cellulose acetate membranes or by absorbing free adenosine on charcoal. The recovery of 1.0–4.0 pmol of adenosine added to blood extracts and carried through purification and radioligand assay averaged 100% with an absolute error of ±0.23 pmol. |
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