EOLA1多克隆抗体的制备及免疫定位Preparation of Rabbit Polyclonal Antibody against EOLA1 and Its Immunolocalization
罗敏,梁自文,杨宗城,罗向东
LUO Min1,LIANG Ziwen2,YANG Zongcheng3,LUO Xiangdong3(1.Cadre Ward
摘要(Abstract):
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 EOLA1,and to explore its immunolocalization in some human malignant tumor tissues.Methods: Recombinant plasmid was constructed 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 EOLA1 in some human malignant tumors was detected with immunohistochemistry.Results: Recombinant plasmid was constructed successfully,and Escherichia coli expressed human EOLA1 protein.Rabbit polyclonal antibody against EOLA1 was obtained.Immunohistochemistry showed that EOLA1 protein expressed in breast carcinoma cell cytoplasm.Conclusions: Anti-EOLA1 polyclonal antibody is successfully prepared,and the expression of EOLA1 protein in breast carcinoma cell cytoplasm is confirmed.
关键词(KeyWords):
EOLA1蛋白;抗体;免疫定位;免疫组织化学
EOLA1 protein;antibody;immunolocalization;immunohistochemistry
基金项目(Foundation): 国家自然科学基金资助项目(30300101,30371468,30670838)
作者(Author):
罗敏,梁自文,杨宗城,罗向东
LUO Min1,LIANG Ziwen2,YANG Zongcheng3,LUO Xiangdong3(1.Cadre Ward
DOI: 10.19367/j.cnki.1000-2707.2012.05.006
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