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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. DOI: 10.11847/zgggws2007-23-03-19
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. DOI: 10.11847/zgggws2007-23-03-19
HE Ping, ZHANG Shi, YIN Yuan-qin, . Preparation and primary purification of rabbit polyclonal antibodies against human Cap43 protein[J]. Chinese Journal of Public Health, 2007, 23(3): 289-291. DOI: 10.11847/zgggws2007-23-03-19
Citation: HE Ping, ZHANG Shi, YIN Yuan-qin, . Preparation and primary purification of rabbit polyclonal antibodies against human Cap43 protein[J]. Chinese Journal of Public Health, 2007, 23(3): 289-291. DOI: 10.11847/zgggws2007-23-03-19

兔抗Cap43多克隆抗体制备和初步纯化

Preparation and primary purification of rabbit polyclonal antibodies against human Cap43 protein

  • 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-硫酸铵法初步纯化抗体后,十二烷基磺酸钠-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.

     

    Abstract: Objective To obtain an anti-Cap43 polyclonal antibody.Methods Peptide with 30 amino acids as hapten was coupled with mcK LH using carbodiimide(EDC)coupling chemical technique to form a coupling ag ent(peptidemcK LH),then rabbits were immunized;To prepare anti-Cap43 polyclonal antibody.Valence of antibody was detected by dot-EL ISA.The polyclonal antiserum was purified by caprylic acid-ammonium sulfate precipitation,then,the purity of antig en was detected by SDS-PAGE.Results Rabbit polyclonal antiserum againsthuman Cap43 protein was obtained after 11 weeks and the titer of the antiserum obtained in this ex periment was up to 1:500.Conclusion Coupling hapten with giant molecular vehicle mcK LH by EDC can make it complete antigen that posessed immunogenicity and immunor eactivity.Immunizing rabbits with the hapten-mcKL H was used to obtain the antibody ag ainst Cap43.

     

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