Genetic studies of human apolipoproteins: XIV. A simple agarose isoelectric focusing gel method for apolipoprotein E phenotyping.
A new and simplified procedure is described for apolipoprotein E (APO E) phenotyping from native plasma or serum samples. Diluted or dialyzed samples are separated on agarose isoelectric focusing gels followed by protein blotting on nitrocellulose membranes. APO E banding patterns are localized immunologically using polyclonal goat anti-APO E antiserum as the primary antibody and rabbit anti-goat IgG conjugated with alkaline phosphatase as the secondary antibody. The method was used in parallel with our previously described polyacrylamide gel system to screen 110 unrelated and healthy US whites. Both gel systems gave identical APO E phenotypes, and allele frequencies were comparable with reported US white values. This simplified method can be used on a large number of population and clinical samples with minimum cost and effort.
Kamboh
M I
MI
Department of Human Genetics, Graduate School of Public Health, University of Pittsburgh, PA 15261.
Kelly
L J
LJ
Ferrell
R E
RE
D001057:Apolipoproteins E / Q000235:genetics*; D005787:Gene Frequency; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D007525:Isoelectric Focusing / Q000379:methods*; D010641:Phenotype; D011110:Polymorphism, Genetic; D014481:United States; D044465:White People
D001057:Apolipoproteins E