Direct Photography of Ultracentrifuge Sedimentation Curves

Philpot, J. St. L.

doi:10.1038/141283a0

Cited by 238 publications

(34 citation statements)

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“…Electrophoretic measurements were made in the Tiselius (1937) apparatus at 00, using a potential gradient of 6v/cm. Optical observations by the diagonal schlieren method (Philpot, 1938) were photographically recorded on Ilford half-tone panchromatic plates, using a high pressure Hg arc as a light source from which monochromatic light 546 mp. was isolated by using a suitable filter.…”

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confidence: 99%

The purification of human fibrinogen

Kekwick

Mackay

Nance

et al. 1955

Biochemical Journal

162

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For investigations concerning the mode of action of the clearing factor, estimations of free fatty acids in mixtures of normal rat plasma and lipaemic dog plasma were performed using a modification of van de Kamer's (1948) method as described for faeces. Because the method developed may be of value for similar or other uses, it is given here. In a 100 ml. flask 2 ml. of the plasma-mixture, containing either heparin or citrate as an anticoagulant, are run into 6 ml. of phosphoric acid solution (metaphosphoric acid, 50 g., NaCl, 250 g. water to 1 1.). After a few minutes 16 ml. of ethanol, containing 0-4 % amyl alcohol, are added, followed by 20 ml. of distilled light petroleum (b.p. 40-60°). The flask is stoppered and the mixture shaken by hand for 1 min. After separation, the upper layer is filtered through dry paper and the funnel covered in order to minimize evaporation. Care should be taken to avoid contamination with traces of the other phase. Ten ml. of the light petroleum extract is evaporated on a boiling-water bath and the residue dried in vacuo. Neutralized ethanol (5 ml.) containing 0-6 mg. of thymol blue per 100 ml. are added and the residue dissolved by boiling gently under reflux for 3 min. Finally, the fatty acids are titrated under a stream of N2 with O-O0N-NaOH.The results are corrected for a blank (usually about 0060 ml. of 001N-NaOH), obtained by running 2 ml. of water through the same procedure.The reproducibility of the method suffices for many purposes. In twenty-four estimations giving results ranging from 021 to 210 0,-equiv. of fatty acids per 2 ml. of plasma tested, the differences between duplicate determinations were found to have a standard deviation of + 0-14 ,u-equiv.

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confidence: 99%

The purification of human fibrinogen

Kekwick

Mackay

Nance

et al. 1955

Biochemical Journal

162

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“…Schlieren diagrams were obtained by the cylindrical lens method (18). Sodium diethyl barbiturate buffer at pH 8.6 and ionic strength 0.1 permitted, in most cases, adequate separation of albumin from alpha-i globulin when the run was carried out at a field strength of approximately 5 volts per centimeter for 2 or 3 hours at 20 to 4°C.…”

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confidence: 99%

Chemical, Clinical, and Immunological Studies on the Products of Human Plasma Fractionation Xxx. The Use of Salt-Poor Concentrated Human Serum Albumin Solution in the Treatment of Chronic Bright's Disease 123

Thorn¹,

Armstrong²,

Davenport³

et al. 1945

J. Clin. Invest.

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Much of the disability induced by chronic Bright's disease, particularly during the nephrotic stage, is attributable to thte coexistent disturbances in protein metabolism. The latter may be characterized as follows: 1, proteinuria; 2, hypoalbuminemia; and 3, depletion of tissue protein stores. During the past few years, numerous attempts have been made to correct the hypoalbuminemia, depletion of body protein stores, and attendant edema in patients with nephrotic syndromes by administering human plasma intravenously (1, 2, 3). This form of therapy obviously had as its objectives the use of protein solution as a diuretic agent by virtue of its high colloid osmotic pressure, the restoration of serum albumin level, and the replenishment of body protein stores. The last two objectives were rarely, if ever, attained because of the limited quantity and high cost of human

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“…During the latter portion of the work, a commercially available Sharples laboratory centrifuge was used. The centrifuge was equipped with a celluloid liner and a cooling co!l, and the experiments wereconductedin a manner similar to that used in work with tobacco mosaic virus (35 (38,39). The solutions used usually contained about 2 to 3 rag.…”

Section: Methodsmentioning

confidence: 99%

An Evaluation of Methods for the Concentration and Purification of Influenza Virus

Stanley¹

1944

Journal of Experimental Medicine

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The concentration and purification of influenza virus by means of differential centrifugation in a vacuum type centrifuge, by adsorption on and elution from adult chicken red cells, by elution of the precipitate formed on freezing and thawing of allantoic fluid, by adsorption on and elution from embryonic chick red cells, and by combinations of the first method with each of the three succeeding methods, have been studied. Over-all yields of virus of about 50 to 70 per cent were obtained by these methods and combinations of methods except for somewhat lower yields when adsorption on and elution from adult chicken red cells was employed. However, the purified products obtained by methods involving only the use of red cells or the freezing and thawing technique were found to contain about 80 per cent of non-virus protein. The purified products obtained when differential centrifugation was used either alone or in combination with any one of the other methods were found to be indistinguishable and to consist of a fairly homogeneous component having a sedimentation constant of about 600 S. Such preparations possessed about 22,000 chicken red cell agglutinating units per mg. of protein nitrogen and solutions containing only about 10–14 gm. of the materials gave 50 per cent infectivity end points in chick embryos. The Sharples centrifuge was found to be almost as efficient as the vacuum type centrifuge for the concentration and purification of influenza virus and, because of its larger capacity, is recommended for the preparation of purified virus on a large scale.

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Direct Photography of Ultracentrifuge Sedimentation Curves

Cited by 238 publications

References 2 publications

The purification of human fibrinogen

The purification of human fibrinogen

Chemical, Clinical, and Immunological Studies on the Products of Human Plasma Fractionation Xxx. The Use of Salt-Poor Concentrated Human Serum Albumin Solution in the Treatment of Chronic Bright's Disease 123

An Evaluation of Methods for the Concentration and Purification of Influenza Virus

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