肝癌相关蛋白HCAP1的表达、抗体制备及其亚细胞定位  

Subcellular location of HCAP1 by its expression and preparation of antiserum

作　　者：王俊茹[1] 覃文新[1] 李锦军[1] 杨艳华[2] 潘勇[1] 万大方[1] 顾健人[1] 

机构地区：[1]上海市肿瘤研究所癌基因及相关基因国家重点实验室,上海200032 [2]中国科学院上海生物工程研究中心,上海200233

出　　处：《中国免疫学杂志》2003年第1期26-28,共3页Chinese Journal of Immunology

基　　金：国家"86 3"生物高技术资助项目 (批准号Z19- 0 2 - 0 1-0 1);国家重点基础研究发展规划"97 3"资金资助项目 (批准号G19980 512 0 9)

摘　　要：目的 :肝癌相关基因HCAP1蛋白产物的表达、抗体的制备及其亚细胞定位。方法 :原核表达HCAP1蛋白 ;纯化的HCAP1蛋白免疫新西兰大白兔 ,制备多抗血清 ;Westernblot分析其在肝癌细胞株的表达 ;通过荧光免疫细胞化学进行亚细胞定位。结果 :获得了较纯的HCAP1表达蛋白和高效价的、高特异性的抗HCAP1的多抗血清 ,并且发现HCAP1大部分位于胞浆 ,少量分布于核仁。结论 :表达的HCAP1蛋白和抗HCAP1的多抗血清可用于HCAP1基因功能的研究 ;HCAP1功能场所可能位于核仁。Objective:To obtain anti HCAP1 polyclonal antiserum and define subcellular localization of HCC associated protein(HCAP1).Methods:The HCAP1 was expressed in E.coli DE3 cells and purified.The antiserum was prepared by immunizing rabbit with purified HCAP1 protein.The expression of HCAP1 was analyzed by Western blot in the HCC cells lines.The subcellular localization was observed using fluoroimmunocytochemistry.Results:The rabbit antiserum was obtained and HCAP1 was localized in cytoplasm and nucleoli.Conclusion:The expressed HCAP1 and anti HCAP1 antiserum could be used for studying the HCAP1 role in the HCC;The expression pattern of HCAP1 was mainly in cytoplasm and it was also detected in the nucleoli.

关 键 词：肝癌 抗体 制备 HCAP1 表达 抗HCAP1多抗 亚细胞定位 

分 类 号：R735.7[医药卫生—肿瘤]