Analytical Ultracentrifugation of Proteins in Solutions of Tyriton X-100 Using Absorption Optics

Abstract

Analytical ultracentrifugation studies on proteins in solutions of the nonionic detergent Triton X-100, by means of a UV scanning system, can be conveniently performed by the following procedure: (1) Scanning of the cells is done at a wavelength of 246 nm. (2) The sedimentation of the detergent is prevented by appropriate adjustment of the solvent density. (3) The reference cell is filled with water. Reliable molecular weight determinations can be performed at detergent concentrations up to 0.4% (w/w) in cells of pathlength d = 12 mm and at correspondingly higher concentrations for d - 3 mm. Approx. 10 ng of protein are sufficient for a determination. The method should be useful for studies of membrane proteins. © 1985, Walter de Gruyter. All rights reserved.