Luley C, Baumstark M W, Wieland H
Zentrallabor der Klinik, Albert-Ludwigs-Universität, Freiburg, Germany.
J Lipid Res. 1991 May;32(5):880-3.
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.
载脂蛋白E异构体的传统测定方法包括对1至5毫升血清进行超速离心、对极低密度脂蛋白（VLDL）进行脱脂，以及在聚丙烯酰胺凝胶中进行等电聚焦（IEF）。为了减少样品体积并避免非特异性蛋白条带，有人提出了免疫印迹法。现在我们描述一种方法变体，该方法使用25微升血清，用聚乙二醇沉淀含载脂蛋白E的脂蛋白替代超速离心，并用去污剂溶解进行脱脂。IEF在琼脂糖中进行。这样每个样品用10微升抗血清就可以对含载脂蛋白E的条带进行特异性免疫固定。该方法产生的载脂蛋白E图谱具有特异性且分辨率良好。此外，它还能大量节省时间和相关设备。
