膜FasL的纯化与诱导凋亡功能的鉴定

Purification and Characterization of Membrane-Bound FasL that Iduces Apoptosis

本文用FasL单抗NOK 1与CNBr Sepharose4B偶联 ,制备亲和层析柱 ,从表达FasL的HEpG2细胞膜蛋白抽提液中分离纯化FasL分子。实验证明用本法纯化的FasL其相对分子质量为 3 10 0 0～ 3 3 0 0 0 ,SDS PAGE呈现单一条带 ,Westernblot表明具有一定的抗原性 ,能诱导表达Fas的细胞发生凋亡 ,在一定浓度范围内呈现量 效关系 ,为进一步研究FasL分子的结构功能及制备抗体提供了实验材料。

In the present study,we purified FasL protein from the crude mixture of HEpG2 cells by affinity chromatography column The purified FasL was shown in SDS gel electrophoresis to have a single band at the position of 31 000～33 000 It's antigenic activity was assayed by Western blotting It could induce apoptosis in Jurkat cells in a dose dependent manner So it is capable of studying the structure and function of FasL and preparing anti FasL antibody