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Atherosclerosis: Experimental Methods and Protocols by Drew A. (ed.)

By Drew A. (ed.)

Kid's medical institution study starting place, Cincinnati, OH. permits biomedical researchers to pick these optimized ideas that could be used to check the improvement, development, and remedy of atherosclerotic lesions. define structure.

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Atherosclerosis: Experimental Methods and Protocols

Kid's health center learn origin, Cincinnati, OH. permits biomedical researchers to pick these optimized options which may be used to review the improvement, development, and therapy of atherosclerotic lesions. define structure.

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Allow the centrifuge to slow over a period of about 4 min from 2000 rpm by using the controlled braking program or by turning off the brake. This allows smooth gradient reorientation. 2. 1. 5 ML) USING THE TLN100 ROTOR IN THE BECKMAN TLX100 (OR OPTIMA 120 OR OPTIMA MAX ULTRACENTIFUGE) 1. 6 mL Liposep. Keep remaining plasma for further analysis. 2. 9 mL) and using a metal cannulae attached to a 2mL syringe layer HEPES-buffered saline on top to fill the tube. 3. Seal the tubes with the plastic stoppers.

4. of Chapter 5). 2 rotors are discussed in Notes 10, 11, and 12, respectively. 4. Notes 1. Although removal of chylomicrons is not strictly necessary if the sole object of the fractionation is to investigate the LDL profile of the plasma, the tendency of the chylomicrons to adhere to the surface of the tube after they have floated up through the gradient means that they can contaminate gradient fractions during collection. 2. Liposep and Maxidens can be purchased from Lipotek Ltd, 34 Meadway, Upton, Wirral, Merseyside, CH49 6JQ, UK.

40 Higgins, Graham, and Davies 3. 1. Preparation of Plasma (see Note 13). 1. Centrifuge the blood at 2000g for 10 min to pellet the leukocytes and erythrocytes. 2. Remove the plasma from the pellet using a plastic Pasteur pipet and transfer it into a plastic vial, setting aside at least 50 ┬ÁL for further analysis. 3. Transfer the plasma to a conical centrifuge tube. 3 mL of HEPES buffered saline on top. 4. Centrifuge the tubes at 13000g for 20 min (see Note 13) to float chylomicrons. 5. Carefully remove the chylomicron-containing layer using an automatic pipet with a wide bore tip.

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