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

By Drew A. (ed.)

Kid's clinic examine beginning, Cincinnati, OH. permits biomedical researchers to choose these optimized innovations that could be used to check the advance, development, and remedy of atherosclerotic lesions. define structure.

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Included also are the preparation of exogenous substrate for CETP-activity assay, isolation of CETP from human plasma, and preparation of CETP–specific polyclonal antibody employed in the ELISA. 2. 1. 1. 1. PREPARATION OF LIPOPROTEIN-DEFICIENT PLASMA (LPDP) 1. 6 mg/mL. Human plasma was collected after centrifugation at 1500g for 10 min at 4°C and stored at 4°C. 2. a. (Nr. 1000, MERCK). 3. MQ-H2O: water prepared by Milli-RO 12 Plus and Milli-Q Plus PF (Millipore). 4. DS: 10 % (w/v) dextran sulfate sodium salt (Code No.

1988) Further resolution of the low density lipoprotein spectrum in normal human plasma: physicochemical characteristics of discrete subspecies isolated by density gradient ultracentrifugation. J. Lipid Res. 29, 442–458. 58 Graham et al. Fig. 2. The cholesterol profiles in iodixanol gradients of peak LDL fractions isolated from KBr gradients (6) from six individuals (A–F). 3. All gradients were unloaded low-density end first by upward displacement with Maxidens (see Note 2). ) measured in a standard spectrophotometric assay.

Transfer 5- and 10-µL aliquots to 96-well microtiter plates, leaving empty wells for standards and blanks. 2. Transfer aliquots of the triglyceride or cholesterol standard to the appropriate wells. 3. Add 200 µL cholesterol or triglyceride assay reagent or follow the manufacturer's instructions. 4. Shake the plates (most plate readers have a plate-shaking facility; alternatively use a flatbed shaker). 5. Allow the color to develop and read the absorbance at 450 nM at 15 and 30 min to ensure the reaction is complete.

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