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小鼠抗人前B细胞白血病同源盒基因3多克隆抗体的制备与鉴定 被引量：1

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摘要 目的制备前B细胞白血病同源盒基因3(PBX3)多克隆抗体。方法采用反转录PCR扩增PBX3基因区序列,并重组入原核表达载体pGEX-4T-1,重组载体转化BL21大肠杆菌,异丙基-β-D硫代半乳糖苷(IPTG)诱导GST-PBX3融合蛋白的表达。SDS-PAGE纯化融合蛋白,并用GST-PBX3融合蛋白免疫BALB/c小鼠,收集经过免疫的小鼠血清即抗PBX3的多克隆抗血清,采用间接ELISA检测其效价,通过免疫细胞化学技术及Western blot法鉴定其特异性。结果成功构建pGEX-4T-1/PBX3原核表达载体,转化BL21菌株后可高效表达GST-PBX3融合蛋白,且以不可溶包涵体形式存在。SDS-PAGE纯化蛋白并免疫小鼠产生的抗PBX3血清可特异识别细胞外源过表达的PBX3蛋白,同时可检测到细胞外源性PBX3蛋白的细胞内定位情况。结论获得效价和特异性良好的PBX3多克隆抗体。
作者 张媛 王力民 赵威 韩海勃
机构地区 北京大学肿瘤医院
出处 《细胞与分子免疫学杂志》 CAS CSCD 北大核心 2014年第11期1176-1179,共4页 Chinese Journal of Cellular and Molecular Immunology
基金 国家自然科学基金(81201964 81372594 81301874 81330051) 北京市自然科学基金(5122012 7132051)
关键词 前B细胞白血病同源盒基因3 融合蛋白表达 多克隆抗体
分类号 R392.11 [医药卫生—免疫学] R392.33 [医药卫生—免疫学]
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参考文献15

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二级参考文献9

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细胞与分子免疫学杂志
2014年 第11期

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