Ultracentrifugal studies in capillary cells. I. Determination of sedimentation coefficients |
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Authors: | J C Pita F J Müller |
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Affiliation: | Department of Medicine, University of Miami School of Medicine, P.O. Box 875, Biscayne Annex, Miami, Florida 33152 USA |
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Abstract: | A technique has been developed which is based on transport method equations and allows determination of sedimentation coefficients using less than 5 ng of a biological active principle. Glass capillaries of 0.30 mm inside diameter and 3.0 mm length are used as sedimentation cells. About 200 nl of pure or impure solutions with concentration as low as 0.05 mg/ml are ultracentrifuged in a swinging-bucket rotor with a conventional preparative ultracentrifuge. The capillary microcells are easily sectioned after centrifugation through a plane previously marked on the glass surface. The technique was successfully extended to the use of larger glass capillaries, up to 1.25 mm inside diameter, and plastic centrifuge tubes of 5.0 mm diameter and 0.40 ml capacity. The method has been experimentally verified with proteins and protein-polysaccharides of known sedimentation constants. |
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