Estimation of lipoprotein and apoprotein distribution in rat plasma by cumulative density ultracentrifugation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis |
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| Authors: | A T H?stmark,E Lystad,A Spydevold ? Y" >Haug |
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| Affiliation: | 1. Institute of Preventive Medicine, University of Oslo, Gydas vei 8, Oslo 3, Norway;2. Institute of Medical Biochemistry, University of Oslo, Sognvannsveien 9 Oslo 3, Norway;1. University of Michigan, Ann Arbor, Michigan;2. Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland;3. Department of Ophthalmology, Massachusetts Eye and Ear, Boston, Massachusetts;4. Harvard Medical School, Boston, Massachusetts;1. Department of Neurosurgery, Inselspital, Bern, Switzerland;2. Department of Neurosurgery, Zurich University Hospital, Switzerland;1. Department of Chemical and Environmental Engineering, The University of Nottingham Ningbo China, Ningbo, 315100, China;2. Key Laboratory of Carbonaceous Wastes Processing and Process Intensification of Zhejiang Province, The University of Nottingham Ningbo China, Ningbo, 315100, China;3. Municipal Key Laboratory of Clean Energy Conversion Technologies of Ningbo, The University of Nottingham Ningbo China, Ningbo, 315100, China;4. Department of Architecture and Built Environment, The University of Nottingham, Nottingham, NG7 2RD, UK;5. Department of Chemical and Environmental Engineering, The University of Nottingham, Nottingham, NG7 2RD, UK;1. Hebei Key Laboratory of Applied Chemistry, School of Environmental and Chemical Engineering, Yanshan University, Qinhuangdao 066004, PR China;2. State Key Laboratory of Metastable Materials Science and Technology, Yanshan University, Qinhuangdao 066004, PR China |
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| Abstract: | Lipoprotein distribution in rat plasma determined after sequential ultracentrifugation (requiring 8 days of centrifugation to separate lipoproteins in five density classes), was compared to estimates based upon cumulative density ultracentrifugation (46 hr of ultracentrifugation). In general comparable values were obtained by the two methods with regard to protein, total cholesterol, cholesteryl ester, free cholesterol, and triacylglycerol distribution. However, the HDL3 protein concentration found by sequential ultracentrifugation was only about 50% of that found after the cumulative procedure. Apolipoproteins in lipoproteins isolated by the two methods were well separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Color of the stained bands was extracted and read photometrically. A linear standard curve was obtained with albumin. Absorbance corresponding to 1 microgram/ml was 0.057. Below d = 1.100 g/ml (HDL2b) the two ultracentrifugation methods gave comparable results for all apoproteins. In contrast to this the level of apo A-I, apo E, and apo A-IV in the more dense types of HDL was higher when estimated by cumulative than by sequential ultracentrifugation. In HDL3 isolated by sequential ultracentrifugation the apo A-IV, apo E, and apo A-I concentrations were 51, 31, and 45% respectively, of values found after cumulative ultracentrifugation. The results indicate that cumulative density ultracentrifugation, followed by colorimetric determination of apoproteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, is a useful approach when studying lipoprotein distribution in rat plasma. |
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