| 兔抗Cap43多克隆抗体制备和初步纯化 |
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| 引用本文： | 何萍,张斯,尹元琴,隋承光,孟凡东,王晓华,张志强,姜又红.兔抗Cap43多克隆抗体制备和初步纯化[J].中国公共卫生,2007,23(3):289-291. |
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| 作者姓名： | 何萍 张斯 尹元琴 隋承光 孟凡东 王晓华 张志强 姜又红 |
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| 作者单位： | 1. 中国医科大学附属第一医院肿瘤研究所,沈阳,110001
2. 哈尔滨市红十字中心医院 |
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| 摘 要： | 目的制备兔抗Cap43多克隆抗体。方法以30个氨基酸组成的多肽为半抗原与钥孔血蓝蛋白（mcKLH）通过碳化二亚胺（EDC）法化学偶联成完全抗原（多肽-mcKLH）免疫家兔；制备兔抗Cap4443多克隆抗体；斑点ELISA法检测抗体效价，经辛酸一硫酸铵法初步纯化抗体后，十二烷基磺酸钠一聚丙烯酰胺凝胶电泳（SDS-PAGE）电泳考马斯亮蓝检测其纯化后的纯度。结果经免疫11周得到兔抗Cap43多克隆体，抗体效价为1：500。结论采用EDC化学偶联半抗原和大分子载体蛋白mcKLH，使其成为同时具有免疫原性与免疫反应性的完全抗原，用其免疫家兔获得多克隆抗体。
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| 关 键 词： | 半抗原 多克隆抗体 斑点ELISA 辛酸-硫酸铵 |
| 文章编号： | 1001-0580(2007)03-0289-03 |
| 修稿时间： | 08 18 2006 12:00AM |
Preparation and primary purification of rabbit polyclonal antibodies against human Cap43 protein |
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| HE Ping, ZHANG Shi, YIN Yuan-qin, et al.Preparation and primary purification of rabbit polyclonal antibodies against human Cap43 protein[J].Chinese Journal of Public Health,2007,23(3):289-291. |
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| Authors: | HE Ping ZHANG Shi YIN Yuan-qin |
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| Affiliation: | Institution of Cancer Research, the Fisrt Affillatied Hospital, China Medical University ,Shenyang 110001, China |
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| Abstract: | Objective To obtain an anti-Cap43 polyclonal antibody.Methods Peptide with 30 amino acids as hapten was coupled with mcKLH using carbodiimide(EDC) coupling chemical technique to form a coupling agent(peptide-mcKLH),then rabbits were immunized;To prepare anti-Cap43 polyclonal antibody.Valence of antibody was detected by dot-ELISA.The polyclonal antiserum was purified by caprylic acid-ammonium sulfate precipitation,then,the purity of antigen was detected by SDS-PAGE.Results Rabbit polyclonal antiserum against human Cap43 protein was obtained after 11 weeks and the titer of the antiserum obtained in this experiment was up to 1:500.Conclusion Coupling hapten with giant molecular vehicle mcKLH by EDC can make it complete antigen that posessed immunogenicity and immunoreactivity.Immunizing rabbits with the hapten-mcKLH was used to obtain the antibody against Cap43. |
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| Keywords: | Cap43 |
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