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| | In Vitro Radioactive (RTA) Assay |
| Description: | An in vitro assay for determining ICso's to identify compounds that inhibit CETP transfer activity is performed based on a modification of a published method (Morton and Zilversmit, (1981) A plasma inhibitor of triglyceride and cholesteryl ester transfer activities, J. Biol. Chem. 256(23), 1 1992-1 1995). The ability of inhibitors to alter CETP activity is performed using two different assays: one using recombinant CETP and one using an endogenous plasma source of CETP. Both assays measure the transfer of [3H] cholesteryl oleate or [3H] triolein from exogenous LDL to HDL.Radiolabeled donor particles are generated by first combining 100 ul of 200 uM butylated hydroxyl toluene in CHC13, 216 of 21.57 mM DOPC in EtOH, and either 500 uCi [3H]-triolein (Perkin Elmer #NET-431) or 500 uCi [3H]-cholesteryl oleate (GE #TRK886) in a glass tube. Reagents are mixed, dried under nitrogen, and then resuspended in 2 mL of 50 mM Tris, 27 uM EDTA. |
| Affinity data for this assay | |
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