Abstract
Publisher Summary Plasma lipoproteins have lower hydrated densities relative to the other plasma proteins; therefore sequential flotation ultracentrifugation has been the principal method used for their isolation and classification. This chapter describes the sequential flotation ultracentrifugation method and some important features for the proper use of sequential flotation ultracentrifugation, such as it is important to calculate the effect of differences in path length or it is virtually impossible to reduce the level of contaminating proteins, such as albumin, below detection limits with a single spin. The ability to process simultaneously large volumes or many small samples is one of the principal advantages of sequential flotation ultracentrifugation. However, the technique suffers from certain limitations, such as during the course of long centrifugation, extensive lipid peroxidation can occur. This problem may be minimized by the inclusion of appropriate chelating agents and antioxidants. Full fractionation and purification by ultracentrifugal methods are both lengthy and costly. Also sequential flotation ultracentrifugation separates lipoproteins according to density only, and the investigator must be aware that particles differing in stage of metabolism, composition, charge, and size may be combined in any of the fractions obtained with this method.
Published Version
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