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基于 Western 印迹技术的基本原理,利用 IgE 结合因子/可溶性 CD23(IgEBF/sCD23)能够和 IgEFC 端特异性结合的特点,在系统研究了包括:IgE 浓度,反应时间及转移条件等数种因素后,首创了 IgE 介导的间接 Western 印迹技术.用于对 SDS—PAGE 电泳分离后样品中 IgEBF/sCD23特异性片段进行鉴定.实践证明:该法具有重复性高,操作简便及节约资金等优点.运用该技术,我们确定了 RPMI8866细胞上清中具有 IgEBF 活性的五种不同分子量的多肽.此方法的建立对开展 IgEBF/sCD23的研究以及其它特异性分子的鉴定都具有参考价值.
Based on the basic principle of Western blotting technique, the specific binding of IgE binding factor / soluble CD23 (IgEBF / sCD23) to IgEFC was studied. After systematically studying several factors such as IgE concentration, reaction time and transfer conditions, The first indirect IgE-mediated Western blotting was used to identify IgEBF / sCD23 specific fragments in SDS-PAGE electrophoresis samples.It has been proved by practice that this method has the advantages of high repeatability, easy operation and cost saving. With this technique, we identified five peptides of different molecular weight with IgEBF activity in the supernatant of RPMI 8866. The establishment of this method is of great value in the research of IgEBF / sCD23 and other specific molecules.