The pro ANF 31-67 radioimmunoassay recognizes in plasma a component of the N-terminus of approximately 3900 mw which is consistent with measuring only aa 31-67 of the prohormone (ie, pro ANF 31-67, mw 3878). Our ANF radioimmunoassay recognizes a 3,000 mw peptide in plasma with ANFs actual molecular weight being 3,081. All determinations were performed in triplicate. The interassay coefficient of variation for pro ANFs 1-30, 31-67, and ANF radioimmunoassays were 4.8 percent, 5.3 percent, and 5.7 percent, respectively. The interassay coefficient of variation was 8 percent for both pro ANFs 1-30 and 31-67 radioimmunoassays while ANFs radioimmunoassay interassay variation was 6.9 percent. buy ortho tri-cyclen
Recovery was examined by adding synthetic unlabeled pro ANF 1-30, pro ANF 31-67 and ANF at 100, 200, and 400 pg/ml to pooled plasma. Recovery for pro ANF 1-30 was 83.5 ± 13.2 (SD) percent while pro ANF 31-67 recovery was 100.9 ±8.9 percent. Recovery of ANF was 92 ±11 percent. The respective IC.*,, were 180, 120 and 11 fmols/tube while the lowest detectable concentrations were 40, 35 and 1.4 fmols for pro ANFs 1-30, 31-67 and ANF radioimmunoassays, respectively. Serial dilution of pooled plasma has revealed excellent parallelism of standard and unknown in these assays. Reverse phase high pressure liquid chromatography utilizing 5 micron cartridge columns revealed that the pro ANFs and ANF measured were authentic.
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