A rapid radiometric assay for adenosine deaminase |
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Authors: | Raymond J. Jackson Richard G. Keightley |
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Affiliation: | 1. Department of Microbiology, University of Alabama in Birmingham, University Station, Birmingham, Alabama, 35294 USA;2. Department of Pediatrics, University of Alabama in Birmingham, University Station, Birmingham, Alabama, 35294 USA |
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Abstract: | A rapid radiochemical procedure for the measurement of adenosine deaminase is described. The method employs phospho-Sephadex, a weak cation exchanger, which permits the enzymic product inosine to pass unretarded through the gel while the radioactive substrate adenosine is retained. Use of a Millipore filter manifold permits rapid processing of samples and eliminates time-consuming column chromatographic, electrophoretic, or paper chromatographic techniques required for separation of product and substrate.The activity of adenosine deaminase was examined in spleen cell preparations prepared from normal CBA mice. Excellent agreement of results was obtained when the radioactive method was compared with two other independent assay techniques. |
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