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小鼠IFIT1多克隆抗体的制备

, PP. 1462-1464

Keywords: IFIT,多克隆抗体,融合蛋白,抗原性,免疫应答

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Abstract:

目的获取小鼠IFIT1(Interferon-inducedproteinwithtetratricopetiderepeats1)特异性多克隆抗体。方法扩增培养高效表达IFIT1的重组子pMAL-C2X-IFIT1,超声破碎后的细胞上清过AmylosePre-packedcolumn亲和层析柱,将所得纯化的融合蛋白MBP-IFIT1作为抗原,添加福氏佐机剂后免疫兔,收集抗血清,用Westernblot和ELISA方法测定抗血清特异性反应和效价。结果纯化的MBP-IFIT1可诱导兔产生特异性免疫应答,所得抗体能够特异性识别融合蛋白中的IFIT1,ELISA结果显示其效价为1∶12800。结论MBP-IFIT1具有良好的抗原性,以该融合蛋白为抗原免疫兔成功制备出IFIT1特异性多克隆抗体。

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