The N-Terminus of the Atrial Natriuretic Factor Prohormone in the Pleural Fluid of Congestive Heart Failure Patients: Patients and Methods (Part 2)

In: Congestive Heart Failure

30 Oct 2012

Pleural Fluids
The mean protein concentration of the six thoracentesis samples was 0.6 g/L with no individual protein concentration above 0.8 g/h in the respective pleural fluids. All of these fluids were, thus, transudates which were handled as follows: 10 ml of each transudate was centrifuged at 3,000 g for 15 minutes and then 100 ml of the supernatant was extracted with 100 percent ethanol (1:1 dilution at 4°C), vortexed, and allowed to stand for 15 minutes. After this 15-minute period, the pleural fluids were re-centrifuged at 3,000 g for 15 minutes and the supernatants taken to dryness with controlled nitrogen flow. The plasma samples (100 m of each) were extracted the same as the pleural fluids. Both plasma and pleural fluids were then examined for their concentration of pro ANF 1-98 (the whole N-terminus of prohormone), pro ANF 31-67 (midportion of N-terminus), and C-terminus (ANF). buy ampicillin
Radioimmunoassays for Pro 1-98, Pro ANF 31-67 and ANF
Radioimmunoassays to measure the N-terminus of the prohormone were devised to aa 1-30 and 31-67 of the 126 aa prohormone while the C-terminal assay measures amino acids 99-126 of the prohormone, ie, ANF, as previously described by our laboratory. Our pro ANF 1-30 radioimmunoassay recognizes a component in plasma of approximately 10,000 mw as characterized by gel-permeation chromatography (G-50 Sephadex) which is consistent with the whole N-terminus of the ANF prohormone (ie, aa 1-98), but without the C-terminus attached to it.

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