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.


About this blog

Blog invites submissions of review articles, reports on clinical techniques, case reports, conference summaries, and articles of opinion pertinent to the control of pain and anxiety in dentistry.