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A Modification Method for Isolation and Acetylation of Low Density Lipoprotein of Human Plasma by Density Discontinuous Gradient Ultracentrifugatio

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Author(s): J.Z. Reza | A. Nikzamir | M. Doosti | M.S. Pour

Journal: Journal of Biological Sciences
ISSN 1727-3048

Volume: 10;
Issue: 8;
Start page: 785;
Date: 2010;
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Keywords: modified low density lipoprotein | Atherosclerosis | lipoproteins | native-low density lipoprotein

ABSTRACT
There is now a consensus that atherosclerosis represents a state of heightened oxidative stress characterized by lipid and protein oxidation in the vascular wall. The purpose of this study was to evaluate a simple and precise manner in which to assess the LDL and modified LDL for clinical and experimental application. A step density gradient was constructed from NaCl and KBr solutions varying in density(d) from 1.035 to 1.1 g mL-1 and from 5.5 mL of pool plasma of 32 healthy normolipedemic men (23-25 years old) was adjusted to d 1.09 g mL-1. The sample was Separated was after a single ultracentrifugation 40,000 g for 10 h at 4°C in a swinging bucket rotor. Centrifugation showed four density layers d = 1.1 g mL-1, d = 1.09 g mL-1, d = 1.065 g mL-1, 1.035 g mL-1. Isolated LDL was acetylated by saturated sodium acetate and acetate anhydride. Acetylated LDL (ac-LDL) has different physico-chemical properties than native LDL. The acetate cellulose electrophoresis was applied for analysis of electrophoretic properties native and ac- LDL. Ac-LDL moved 1.5 times faster than native LDL. This study support a simple and precise manner in which to assess the LDL and modified LDL (oxidized -LDL and acetylated-LDL) for clinical and research purposes.
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