Havekes L M, de Knijff P, Beisiegel U, Havinga J, Smit M, Klasen E
J Lipid Res. 1987 Apr;28(4):455-63.
A new method for the apolipoprotein E phenotyping has been developed. The method is based on isoelectric focusing of either delipidated or guanidine-HC1-treated serum or plasma in a horizontal slab gel system followed by immunoblotting using either polyclonal or monoclonal anti-apolipoprotein E antibodies as first antibody. Apolipoprotein E phenotyping with this method in 200 serum samples that had been stored at -20 degrees C for more than one year gave exactly the same results as obtained with the conventional method based on isoelectric focusing of delipidated very low density lipoproteins isolated from fresh serum followed by protein staining. Compared with the conventional method, the present method is less laborious because ultracentrifugation to isolate VLDL is not needed; it is suitable for large scale screening purposes; it needs only a few microliters of serum or plasma, and can easily be performed with samples with low concentrations of apolipoprotein E.
一种新的载脂蛋白E表型分析方法已被开发出来。该方法基于在水平平板凝胶系统中对脱脂或经盐酸胍处理的血清或血浆进行等电聚焦，然后使用多克隆或单克隆抗载脂蛋白E抗体作为一抗进行免疫印迹。用这种方法对200份在-20℃下储存了一年以上的血清样本进行载脂蛋白E表型分析，其结果与基于对从新鲜血清中分离的脱脂极低密度脂蛋白进行等电聚焦然后进行蛋白质染色的传统方法所得到的结果完全相同。与传统方法相比，本方法工作量较小，因为不需要超速离心来分离极低密度脂蛋白；它适用于大规模筛查；只需要几微升血清或血浆，并且可以很容易地对载脂蛋白E浓度低的样本进行检测。
