Human Isoionic Hemoglobin: Preparation and Kinetic PropertiesBerger, Robert L.; Carpenter, Lowell; Everse, Johannes; Kaplan, Jerold L.
doi: 10.1080/00032717308062188pmid: N/A
Abstract Much of the data on hemoglobin-gas interactions which have accumulated in the literature is difficult to interpret because the hemoglobin preparations contained extraneous ligands and salts. In order to obtain hemoglobin which is completely free of 2, 3-DPG and other ionic substances, we have developed a method for isolating isoionic hemoglobin. These data serve as a baseline against which other data obtained with added ligands and salts can be compared. Specifically, isoionic hemoglobin had these properties: Second-order rate constant for combination with 02: 2.2 × 107 M−1S−1; p50 at 20[ddot] was 0.87mm Hg at heme concentration of 10−4 M.
The Measurement of Nanogram Quantities of Vitamin D3Campion, T. H.; Dilley, Susan
doi: 10.1080/00032717308062189pmid: N/A
Abstract An isotope dilution method is described for the measurement of nanogram quantities of vitamin D3 (cholecalciferol). Use is made of the Diels -Alder reaction between vitamin D3 and tetracyanoethylene. Increasing quantities of exogeneous vitamin D3 added to a standard reaction mixture of 14C-labelled vitamin D3 and tetracyanoethylene produced a decrease in the ratio of reacted to unreacted vitamin D3. The ratio (y) was measured by radio-scanning of an eluted thin-layer chromatogram, and quantitation of added vitamin D3 was thereby achieved.
Radioimmunoassay of Plasma Pregnenolone-SulfateBuster, John E.; Abraham, Guy E.
doi: 10.1080/00032717308062190pmid: N/A
Abstract This report describes a radioimmunoassay method for the direct measurement of pregnenolone-sulfate (5Δ-P-S) in diluted plasma. Utilizing a specific anti-5Δ-P-S antiserum which reacted both with pregneolone (5Δ-P) and its sulfate (5Δ-P-S), we have measured 5Δ-P-S with accuracy in small aliquots of diluted plasma. Because the plasma concentration of 5Δ-P-S is relatively high, the effect of 5Δ-P and other steroids on the assay was negligible. The usable range of the standard curve was between 0.1 and 25 ng of 5Δ-P-S. The range of values obtained by this method for plasma 5Δ-P-S in peripheral maternal plasma and in the newborn compared favorably with the range of levels observed with other reliable but more cumbersome methods.
Separation of Prostaglandins on a Hydrophobic Sephadex DerivativeNyström, Ernst; Sjövall, Jan
doi: 10.1080/00032717308062191pmid: N/A
Abstract Liquid-gel chromatography on a hydrophobic Sephadex derivative has been used for the separation of prostaglandins. Solvent systems are given for separation of PGE and PGF compounds either as free acids or as methyl esters, using a Sephadex derivative containing 12% hydroxyalkyl groups. The columns are simple to prepare and can be used repeatedly over long periods of time. The recovery is high and the technique should be a useful complement to other methods used for the isolation of prostaglandins.
Isothermal Titration CalorimeterHawkes, C. M.; Janata, J.
doi: 10.1080/00032717308062192pmid: N/A
Abstract The construction and performance of an isothermal titration calorimeter is described. It is based on continuous automatic balancing of constant Peltier cooling with variable Joule heating. By using this arrangement the short response time (∼5 s) obtained, makes this instrument particularly suitable for applications to solution kinetics and for thermometric titrations.