Rapid apolipoprotein E phenotyping by immunofixation in agarose.
Conventional determination of apolipoprotein E isomorphs comprises ultracentrifugation of 1-5 ml serum, delipidation of very low density lipoproteins (VLDL), and isoelectric focusing (IEF) in polyacrylamide gels. In order to reduce the sample volume and to avoid nonspecific protein bands, immunoblotting was proposed. Now we describe a methodological variant that uses 25 microliters serum, replaces ultracentrifugation by precipitation of apoE-containing lipoproteins with polyethylene glycol, and delipidation by dissolution in detergent. IEF is carried out in agarose. This allows specific immunofixation of apoE-containing bands with 10 microliters antiserum per sample. This method yields apoE patterns that are specific and well resolved. Also, it offers considerable savings of time and equipment involved.
Luley
C
C
Zentrallabor der Klinik, Albert-Ludwigs-Universität, Freiburg, Germany.
Baumstark
M W
MW
Wieland
H
H
D001057:Apolipoproteins E / Q000032:analysis* / Q000145:classification; D011232:Chemical Precipitation; D006801:Humans; D007158:Immunologic Techniques; D007525:Isoelectric Focusing / Q000379:methods; D010641:Phenotype; D012685:Sepharose
D001057:Apolipoproteins E; D012685:Sepharose