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-  2017 

抗鼠跨膜型TNF-α多克隆抗体设计和制备
Design and preparation of a polyclonal antibody against TNF-alpha in rats

DOI: 10.11778/j.jdxb.2017.01.005

Keywords: 跨膜型TNF-α,多克隆抗体,酶联免疫吸附试验,流式细胞术
transmembrane TNF-α
,polyclonal antibody,enzyme linked immunosorbent assay,flow cytometer technique

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Abstract:

摘要 目的设计和制备区分鼠跨膜型肿瘤坏死因子-α(TNF-α)和分泌型TNF-α的多克隆抗体.方法: 采用抗原肽预测软件对跨膜型TNF-α表位进行预测,合成跨膜型TNF-α24个氨基酸多肽,并与匙孔碱血蓝蛋白偶联,家兔皮背部多点注射,收集分离血清,采用酶联免疫吸附试验和流式细胞术鉴定其特异性.结果: 制备出高效价的抗血清,该抗血清可特异性结合免疫原多肽,不与分泌型TNF-α发生交叉反应,流式细胞术显示该抗血清可与细胞系NIH 3T3 表达的跨膜型TNF-α发生特异性结合.结论: 成功制备了跨膜型TNF-α特异性多克隆抗体,为区分跨膜型TNF-α和分泌型TNF-α生物学作用提供了新工具.

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