Lipoprotein Protocols

About this book

Jose Ordovas compiles a cutting-edge collection of molecular protein techniques for studying advanced aspects of lipoprotein structure and metabolism. These step-by-step methods allow both the beginning and the advanced researcher to carry out not only fundamental, but also the most advanced genetic screening techniques, including RT-PCR and the preparation of relevant models of transgenic and knockout mice. In addition, novel techniques are presented to characterize plasma lipoproteins, their subfractions, and their modifications, and to determine the activity of relevant enzymes and carrier proteins. Each method is described in easily reproducible detail by a hands-on expert who has perfected it. "Lipoprotein Protocols" offers many powerful tools for unraveling the complexity of lipoprotein metabolism and determining the individual risk of coronary heart disease.

Contents

Part I: dna and rna Based Methodology.PCR Methodology Applied to Genetic Studies of Lipoprotein Metabolism and AtherosclerosisShelley A. Cole and James E. Hixson. Separation of Small-Size DNA Fragments Using Agarose Gel ElectrophoresisJose M. Ordovas. Quantification of mRNA by Polymerase Chain Reaction (PCR) Using an Internal Standard and a Nonradioactive Detection MethodWilliam H. Karge III Ernst J. Schaefer and Jose M. Ordovas. Generation of Transgenic Mice Marten H. Hofker and Marco Breuer. Preparation of Knockout Mice JesGBPs Osada and Nobuyo Maeda. Part II.Protein Based Methodology.Fast Ultracentrifugation Methods for the Separation of Plasma LipoproteinsJose M. Ordovas. Preparative Isolation of Plasma Lipoproteins Using Fast Protein Liquid ChromatographyJose M. Ordovas and Doreen Osgood. Separation of Polipoproteins by Polyacrylamide Gel ElectrophoresisJose M. Ordovas. Isolation and Purification of Serum Lipoprotrein (a) Using Lectin AffinityLeo J. Seman Carl de Luca and Jennifer L. Jenner. Electrophoretic Separation of LDL and HDL Subclasses David L. Rainwater. Measurement of Lipoprotein ParticlesCatherine Fievet and Jean-Charles Fruchart. Methodological Approaches for Assessing Lipid and Protein Oxidation and Modification in Plasma and Isolated LipoproteinsWolfgang Sattler Ernst Malle and Gert M. Kostner. Separation and Quantitation of Phospholipid Classes by HPLCGeorge M. Patton and Sander J. Robins. Assays of Lecithin Cholesterol Acyltransferase (LCAT) Milada Dobi sov and Jiri J. Frohlich. Determination of the Mass Concentration and the Activity of the Plasma Cholesteryl Ester Transfer Protein (CETP)Laurent Lagrost. In Vitro Measurement of Lipoprotein and Hepatic LipasesElisabet Vilella and Jorge Joven. Cellular Assays for Lipoprotein ReceptorsSusan Acton and Attilio Rigotti . Measurements of Proteoglycan-Lipoprotein Interaction by Gel Mobility Shift AssayEva Hurt-Camejo Germ n Camejo and Peter Sartipy.

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