中国实验动物学报2012，Vol.20Issue(1)：84-87,4.DOI:10.3969/j.issn.1005-4847.2012.01.018
兔抗金黄地鼠酶标抗体的制备及应用
Preparation and application of the rabbit anti-hamster IgG-HRP
摘要
Abstract
Objective To purify IgG from hamster serum, and to prepare the rabbit anti-hamster IgG-HRP, to be used for detection of the antibody from Sendai virus ( SV)-infected hamsters. Methods Using HiTrap Protein-A affinity chromatography column to purify the IgG from hamster serum. The purity of the hamster-IgG was identified by SDS-PAGE analysis. To produce the hamster against rabbit IgG, and to measure the titer of rabbit anti-hamster IgG by double immunodiffusion. To purify the rabbit anti-hamster IgG by affinity chromatography and prepare the rabbit anti-hamster HRP-con-jugated antibodies. The working concentration of the rabbit anti-hamster IgG-HRP was detected by direct ELISA and Western blot, and detect the antibody from Sendai virus-infected hamster by enzyme immunoassay (IEA). Results The hamster IgG was purified up to 95% determined by SDS-PAGE analysis, and the titer of the antisrum was up to 1:64 assessed by double immunodifusion. The working concentration of the rabbit anti-hamster IgG was up to 1 ;5000 measured by direct ELISA, and 1 :2000 measured by Western blot. The IEA titer was 1:2000. Conclusions The method has been established for efficient purification of IgG fraction from hamster serum, and the polyclonal antibody of the hamster-IgG and the rabbit anti-hamster IgG-HRP are prepared, which may provide a basis for development of serological test system of microbi-al pathogens of hamster.关键词
金黄地鼠/IgG纯化/酶标抗体Key words
Golden-hamster/IgG/Affinity chromatography/Rabbit anti-hamster IgG-HRP分类
生物科学引用本文复制引用
刘先菊,林树柱,杨帆,佟巍,刘云波..兔抗金黄地鼠酶标抗体的制备及应用[J].中国实验动物学报,2012,20(1):84-87,4.基金项目
中央级公共性科研院所基本科研业务费专项资金项目(项目编号:DWS200906). （项目编号:DWS200906）
