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Clinical Chemistry 34: 2275-2279, 1988;
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Clinical Chemistry, Vol 34, 2275-2279, Copyright © 1988 by American Association for Clinical Chemistry

A highly specific radioreceptor assay for the active circulating form of atrial natriuretic factor in human plasma

H Ong, A De Lean and C Gagnon
Clinical Research Institute, Montreal, Quebec, Canada.

This highly sensitive radioreceptor assay (RRA) for the active circulating form of atrial natriuretic factor (ANF), fragment 99-126, in human plasma requires 125I-labeled ANF, bovine zona-glomerulosa membrane receptors, and amiloride HCl. The amiloride elicits an increase in the binding affinity of ANF to its receptors. ANF is extracted from human plasma with "Sep-Pak" cyano cartridges. After a 90- min incubation at 25 degrees C, bound and free fractions are separated by filtration. The ANF concentration that inhibits receptor binding of the radioligand by 50% is 12.4 fmol of unlabeled ANF per tube. The minimum detectable concentration is 0.2 fmol per tube. Using ANF- supplemented plasma samples, there was a good correlation (r = 0.99) between ANF concentrations found and those expected. Only the active circulating form of ANF fully cross-reacts in this assay, which confirms its high selectivity. Results correlated strongly (r = 0.93) for clinical samples tested by RRA and RIA.




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Copyright © 1988 by the American Association for Clinical Chemistry.