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| Preparation and application of the rabbit anti-hamster IgG-HRP |
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| DOI：10.3969/j.issn.1005-4847.2012.01.018 |
| KeyWord:Golden-hamster； IgG； Affinity chromatography； Rabbit anti-hamster IgG-HRP |
| Author | Institution |
| 刘先菊 |
中国医学科学院实验动物研究所 卫生部实验动物检测中心, 北京 |
| 林树柱 |
中国医学科学院实验动物研究所 卫生部实验动物检测中心, 北京 |
| 杨帆 |
中国医学科学院实验动物研究所 卫生部实验动物检测中心, 北京 |
| 佟巍 |
中国医学科学院实验动物研究所 卫生部实验动物检测中心, 北京 |
| 刘云波 |
中国医学科学院实验动物研究所 卫生部实验动物检测中心, 北京 |
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| Abstract: |
| ObjectiveTo 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. MethodsUsing 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-conjugated 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). ResultsThe 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. ConclusionsThe 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 microbial pathogens of hamster. |
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