摘要
目的分离纯化黄鳝血清免疫球蛋白,制备其兔抗血清,并检测抗血清的特异性。方法用Protein A亲和层析的方法纯化黄鳝血清免疫球蛋白,通过SDS-聚丙烯酰胺凝胶电泳检测蛋白的纯度,免疫大耳白兔制备抗血清,利用免疫双扩散检测抗血清的效价,通过western blotting检测抗血清的特异性。结果纯化了黄鳝血清免疫球蛋白,免疫双扩散法测定兔抗黄鳝免疫球蛋白血清效价为1∶32,western blotting结果显示抗血清具有很好的特异性。结论成功纯化了黄鳝免疫球蛋白,制备了兔抗黄鳝IgM抗血清,为建立黄鳝的血清学检测系统奠定了基础。
Objective To purify the immunoglobulin from Ricefield eel, and to prepare the rabbit polyclonal antisera. Methods Ricefield eel immunoglobulin was purified using staphylococcal protein A affinity chromatography. The products were examined by SDS-PAGE electrophoresis. Rabbit antisera were prepared by repeatedly immunizing rabbits with purified IgM. And anti-Ricefield eel IgM antibodies in rabbit serum were detected by agarose double immunodiffusion. Western blotting was performed to assess specificity of rabbit antisera for Ricefield eel IgM. Results Immunoglobulin purified from l^icefield eel had only two bands in SDS-PAGE. The titer of rabbit antisera obtained was up to 1 ： 32 examined by agarose double immunodiffusion. The specific activity of polyclonal rabbit antisera against immunoglobulin of Ricefield eel was indicated with western blotting. Conclusions Ricefield eel immunoglobulin was successfully purified, and rabbit antisera was prepared. This provide a tool for developing the serological test system of Ricefield eel.
出处
《中国比较医学杂志》
CAS
2007年第11期644-647,共4页
Chinese Journal of Comparative Medicine
基金
社会公益研究专项(2005DIA1J095)
关键词
亲和层析
黄鳝
免疫球蛋白
抗血清
Affinity chromatography
Ricefield eel
Immunoglobulin
Antisera
