Quantification of apolipoproteins B-100, B-48, and E in human triglyceride-rich lipoproteins.
We have developed a procedure to quantify apolipoprotein (apo) B-100, apoB-48, and apoE in human triglyceride-rich lipoproteins. This procedure permits delipidation of small amounts of triglyceride-rich lipoproteins without appreciable losses, and quantification of these apolipoproteins in samples containing as little as 10 micrograms of protein. Delipidated triglyceride-rich lipoproteins are subjected to sodium dodecyl sulfate polyacrylamide slab gel electrophoresis, and the mass of apolipoproteins is estimated after densitometric scanning and volume integration of Coomassie blue-stained bands. The chromogenicities of apoB-100 and apoB-48 are virtually identical, and twofold lower than that of apoE. The standard curve for each apolipoprotein follows a power function over a wide protein range, permitting quantification of as little as 0.2 microgram of apoB-48 and as much as 30 micrograms of apoB-100 from a single application of triglyceride-rich lipoproteins to the gels. This method is suitable for routine use in studies of the intestinal and hepatic contributions to triglyceride-rich lipoproteins and their responses to postprandial lipemia.
Kotite
L
L
Cardiovascular Research Institute, University of California, San Francisco 94143-0130, USA.
Bergeron
N
N
Havel
R J
RJ
D053299:Apolipoprotein B-100; D053283:Apolipoprotein B-48; D001055:Apolipoproteins B / Q000032:analysis*; D001057:Apolipoproteins E / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008074:Lipoproteins / Q000737:chemistry*; D014280:Triglycerides / Q000032:analysis / Q000737:chemistry*
D053299:Apolipoprotein B-100; D053283:Apolipoprotein B-48; D001055:Apolipoproteins B; D001057:Apolipoproteins E; D008074:Lipoproteins; D014280:Triglycerides