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作 者:罗敏[1] 梁自文[2] 杨宗城[3] 罗向东[3]
机构地区:[1]中国人民解放军第44医院干部病房,贵州贵阳550009 [2]第三军医大学西南医院内分泌科,重庆400038 [3]第三军医大学西南医院烧伤研究所,重庆400038
出 处:《贵阳医学院学报》2012年第5期476-479,482,共5页Journal of Guiyang Medical College
基 金:国家自然科学基金资助项目(30300101;30371468;30670838)
摘 要:目的:制备兔抗人内皮高表达脂多糖相关因子1(EOLA1)多克隆抗体,探讨其在部分人体恶性肿瘤组织中的免疫定位。方法:构建重组质粒,转化大肠杆菌及诱导表达,抽提目的蛋白免疫家兔制备多克隆抗体,免疫组织化学法检测EOLA1在部分人体恶性肿瘤组织中的表达。结果:成功制备重组质粒在大肠杆菌中表达人EOLA1蛋白,免疫家兔制备得到兔抗EOLA1多克隆抗体,免疫组织化学法检测显示EOLA1蛋白在乳腺癌组织癌细胞胞浆中有表达。结论:成功制备EOLA1多克隆抗体,证实EOLA1蛋白在乳腺癌组织癌细胞胞浆中有表达。Objective: To prepare rabbit polyclonal antibody against EOLAI, and to explore its im- munolocalization in some human malignant tumor tissues. Methods: Recombinant plasmid was con- structed and transformed into Escherichia coli to induce the expression of target protein. Target protein was extracted to immunize rabbits to prepare polyclonal antibody. The expression of EOLAI in some human malignant tumors was detected with immunohistochemistry. Results: Recombinant plasmid was constructed successfully, and Escherichia coli expressed human EOLA1 protein. Rabbit polyclonal an- tibody against EOLA1 was obtained. Immunohistochemistry showed that EOLA1 protein expressed in breast carcinoma cell cytoplasm. Conclusions: Anti-EOLAI polyclonal antibody is successfully pre- pared, and the expression of EOLA1 protein in breast carcinoma cell cytoplasm is confirmed.
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