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CETP Activity Assay Kit II (Fluorometric) (ab196995) uses a donor molecule containing a fluorescent self-quenched phospholipid that is transferred to an acceptor molecule in the presence of Cholesteryl Ester Transfer Protein (CETP). CETP-mediated transfer of the fluorescent phospholipid to the acceptor molecule results in an increase in fluorescence (Ex/Em = 480/511 nm). The fluorometric intensity is directly proportional to the amount of phospholipid transferred. The kit contains rabbit serum and torcetrapib (CETP inhibitor) as positive and negative control, respectively, for assay validation.
This assay can measure PLTP activity in plasma and serum but can also be used for testing activity of recombinant CETP protein.
Cholesteryl Ester Transfer Protein (CETP) is a member of the lipid transfer/lipopolysaccharide binding protein gene family. CETP is plasma protein that transfers a cholesteryl ester from HDL to LDL or VLDL in exchange for a triglyceride. HDL plays an important role in lipid metabolism and cardiovascular health. HDL transports cholesterol to the liver for excretion or to steroidogenic tissues for steroid synthesis. HDL also plays an important role in the reverse cholesterol transport pathway, removing cholesterol from lipid-filled macrophages, protecting against atherosclerosis. Because of this function, CETP is viewed as a target to increase HDL, with CETP inhibition an active area of research and several CETP inhibitors at various stages of drug development.
|CETP Acceptor Molecule||1 x 0.5ml|
|CETP Assay Buffer||1 x 20ml|
|CETP Inhibitor||1 x 10µl|
|CETP Positive Control||1 x 0.1ml|
|Donor Molecule||1 x 0.5ml|
Typical Donor molecule Standard Curve obtained following assay protocol.
Measurement of CETP activity on 1 µL rabbit serum (dark blue diamonds), 1 µL human plasma (orange squares) or 800 ng recombinant human CETP (purple circles). Background control with no serum is shown (green triangles).
Inhibition curve of CETP activity from rabbit serum by Torcetrapib (CETP inhibitor). The assay was run for 1 hour and the IC50 was determined to be 3.56 nM.
Inhibition of CETP activity from human plasma using 80 nM Torcetrapib; assay was run for 2 hours following protocol instructions.
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