Rapid preparation of low density lipoprotein from normal rabbit serum.
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概要
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A rapid method for the isolation of pure low density lipoprotein (LDL) from normal rabbit serum within 6.5h was developed by a combination of discontinuous density gradient ultracentrifugation in a vertical rotor and Fast Protein, Polypeptide, Polynucleotide Liquid Chromatography (FPLC) system with a Superose 6 column. Analyses by FPLC, agarose gel electrophoresis, and sodium dodecyl sulfatepolyacrylamide gel electrophoresis showed that LDL isolated by the present method is free from the contamination of other lipoproteins and serum proteins, while that by the standard method [Hatch, F.T., and Lees, R.S. (1968) Adv. Lipid Res., 6, 1-68] contained very low density lipoprotein (VLDL) and small amounts of proteins such as albumin. Differing from the case of the standard method, significant lipid peroxidation of the sample does not seem to occur in the present method. This method can also be used for the rapid preparation of VLDL and high density lipoprotein.
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