März W, Cezanne S, Gross W
Gustav Embden-Center of Biological Chemistry, J. W. Goethe-University, Frankfurt/Main.
Electrophoresis. 1991 Jan;12(1):59-63. doi: 10.1002/elps.1150120111.
An immunoblotting method for the determination of apolipoprotein E (apoE) phenotypes has been developed. Delipidated plasma proteins are focused in an immobilized pH gradient, and transferred to polyvinylidene difluoride (PVDF) membranes. ApoE isomorphs are identified by immunoperoxidase staining. The method allows reproducible assignment of apoE phenotypes without isolation of triglyceride-rich lipoproteins. Only small amounts of serum are required. There are several important steps in the procedure: (i) delipidation is indispensable, (ii) carrier ampholytes have to be added to the gels and to the sample buffer, and, (iii) on immunostaining, polyvinylidene difluoride membranes provide an excellent signal-to-background ratio.
已开发出一种用于测定载脂蛋白E（apoE）表型的免疫印迹方法。脱脂血浆蛋白在固定化pH梯度中聚焦，然后转移到聚偏二氟乙烯（PVDF）膜上。通过免疫过氧化物酶染色鉴定apoE异构体。该方法无需分离富含甘油三酯的脂蛋白即可对apoE表型进行可重复的鉴定。仅需少量血清。该过程有几个重要步骤：（i）脱脂必不可少，（ii）必须向凝胶和样品缓冲液中添加载体两性电解质，以及（iii）在免疫染色时，聚偏二氟乙烯膜可提供出色的信号与背景比。
