| 吴晋元 周艳 解裕萍 唐东红 叶尤松 李鸿钧△ 孙茂盛△.树鼩IgG纯化鉴定及其多克隆抗体制备和检测[J].,2016,16(7):1201-1204 |
| 树鼩IgG纯化鉴定及其多克隆抗体制备和检测 |
| Purification of Tree Shrews (Tupaia) Immunoglobulin G and Preparation ofAnti- IgG Polyclonal Antibody |
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| DOI： |
| 中文关键词: 树鼩 免疫球蛋白 纯化 多克隆抗体 免疫印记 |
| 英文关键词: Tree shrews Immunoglobulin Purification Polyclonal antibody Western Blot |
| 基金项目:国家科技支撑计划" 实验用树鼩的标准化研究和人类重大疾病树鼩模型创建与应用集成示范" 项目（2014BAI01B01~01） |
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| 中文摘要: |
| 目的：比较两种抗体纯化方法在分离纯化树鼩IgG抗体的应用，制备抗IgG 的多克隆抗体及检测。方法：采用两种商品化
IgG 抗体纯化试剂盒分离树鼩血清IgG 抗体，采用SDS-PAGE 和蛋白定量测定提纯IgG。以树鼩IgG作为抗原，与等量弗氏完全
佐剂（第一次）、弗氏不完全佐剂（第二次）混合皮下注射免疫兔，对分离血清进行多克隆抗体纯化及Western Blot 检测及定量分
析。结果：两种方法均能有效分离纯化树鼩IgG，在经过Montage PROSEP-A试剂纯化后的IgG 在纯度和含量方面均优于Protein
A/G Matrix 试剂。通过纯化后的树鼩IgG免疫兔制备的抗IgG 抗体能有效识别树鼩IgG。结论：纯化的树鼩IgG 具有良好免疫原
性，由此制备的抗体具有高度特异性。研究结果为利用树鼩作为实验动物提供了必要的实验基础。 |
| 英文摘要: |
| Objective:To isolate and purify tree shrew IgG antibody, and perform the preparation and detection of anti-IgG polyclonal
antibody.Methods:The two commercial IgG antibody purification kits were used to separate tree shrew serum IgG, determination
of purified IgG by SDS-PAGE and protein quantification were also performed. The rabbits were immune injected with tree shrew IgG as
antigen, combined with the equivalence Freund's complete adjuvant as prime, incomplete Freund's adjuvant as boost. Western Blot was
performed to detect and analyze the purification and preparation of polyclonal antibody.Results:The two purification methods could effectively
separate and purify the tree shrew IgG, the purity and content of IgG after purification of Montage PROSEP-A reagent were better
than using Protein A/G Matrix kit. The anti-IgG antibodies after purification against tree shrew IgG immunization with rabbit could effectively
identify the tree shrew IgG.Conclusion:the purification of tree shrew IgG had good immunogenicity, thus prepared with a
highly specific antibody. The results supported for usage of tree shrews as an experimental animal to providing experimental basis. |
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