Ultracentrifugation micromethod for preparation of small experimental animal lipoproteins

Analytical Biochemistry
Víctor Rodríguez-SuredaJulia Peinado-Onsurbe

Abstract

Sequential flotation ultracentrifugation is commonly used in the preparation of plasma lipoproteins. However, protocols often require prolonged centrifugation time (48-72 h) and large plasma volumes (2-20 ml), which makes them unsuitable for studies on small laboratory animals. Although analytical techniques such as FPLC have often small sample requirements, further fraction analysis is often limited to the small fraction volume obtained. A sequential ultracentrifugation micromethod is described to obtain rat lipoprotein fractions from 400 microl of plasma in a cumulative centrifugation time of 7.5 h. Fraction volumes were determined and densities were adjusted to those of rat plasma lipoproteins. Polyacrylamide gel electrophoresis and enzymatic measurements of triglycerides, total cholesterol, and phospholipids were used to assess the purity of the lipoprotein fractions. The results were compared with those obtained from a classical sequential ultracentrifugation protocol. The micromethod presented here can be further adapted to other experimental animal species with little modifications.

References

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Citations

Feb 19, 2009·Toxicological Sciences : an Official Journal of the Society of Toxicology·Ezra R LoweMichael J Bartels
May 27, 2008·Journal of Neuroscience Research·Carolina ParadaDavid Bueno
Sep 3, 2003·Journal of Lipid Research·Xavier GalanIgnasi Ramírez

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