Genetic studies of human apolipoproteins: XIV. A simple agarose isoelectric focusing gel method for apolipoprotein E phenotyping.
Genetic studies of human apolipoproteins: XIV. A simple agarose isoelectric focusing gel method for apolipoprotein E phenotyping.
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人类载脂蛋白的遗传学研究：XIV。
DOI：
10.1002/elps.1150110408
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发表时间：
1990
期刊：
影响因子：
2.9
通讯作者：
Ferrell,RE
中科院分区：
文献类型：
--
作者：
Kamboh,MI;Kelly,LJ;Ferrell,RE
描述了一种新的简化程序，用于从天然血浆或血清样本中进行载脂蛋白 E (APO E) 表型分析。稀释或透析的样品在琼脂糖等电聚焦凝胶上分离，然后在硝酸纤维素膜上进行蛋白质印迹。使用多克隆山羊抗 APO E 抗血清作为一抗，使用与碱性磷酸酶缀合的兔抗山羊 IgG 作为二抗，对 APO E 条带模式进行免疫学定位。该方法与我们之前描述的聚丙烯酰胺凝胶系统并行使用，以筛选 110 名不相关且健康的美国白人。两种凝胶系统均给出相同的 APO E 表型，且等位基因频率与报道的美国白色值相当。这种简化的方法可以以最小的成本和工作量用于大量人群和临床样本。
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.