人完整p53凋亡刺激蛋白家族抑制成员多克隆抗体的制备及鉴定  

Preparation and identification of polyclonal antibody of human full-length iASPP

作　　者：蔡云[1] 高兴[1] 辛海明[1] 陈杰[1] 扬新[1] 刘泽军[1] 

机构地区：[1]第三军医大学西南医院国际合作实验室,重庆400038

出　　处：《医学研究生学报》2009年第3期240-243,共4页Journal of Medical Postgraduates

基　　金：重庆市科委自然科学基金资助项目(批准号:2008BA5003)

摘　　要：目的:p53凋亡刺激蛋白(ASPP)家族可调节p53抑癌功能,其抑制成员(iASPP)具有特异性抑制p53诱导细胞凋亡的能力。为了对iASPP进行系统研究,文中报道制备完整的iASPP分子多克隆抗体的方法。方法:在完整iASPP独有的N端选择3个免疫原肽段,制备了3个多克隆抗体。用ELISA检测抗体效价,Western blot比较3个抗体检测内源性抗原的效果,并用体外转录翻译系统制备的完整iASPP蛋白作为对照标准。结果:1号抗体效价>1∶32000,2号和3号抗体效价>1∶16000。Western blot确定100000为完整iASPP蛋白的条带位置。结论:成功地制备了iASPP的多克隆抗体,为深入研究完整iASPP提供了有用的工具。Objective：Apoptosis stimulating protein of p53 （ASPP） may regulate the inhibitory effects on tumor induced by p53. The inhibitory member of ASPP family （iASPP） is capable of specially inhibiting p53 induced apoptosis. In this study a polyclonal anti-iASPP antibody specific to whole iASPP was prepared to investigate the function of iASPP. Methods ： We generated antibodies using different antigens contained sequence specific to N-terminus of iASPP. Valence of antibody was detected by ELISA. Western blots were used to compare the efficiency of three antibodies in extracts from a range of cell lines derived from different tissues. Results：The iASPP in vitro translated was used as a marker to confirm the band position of iASPP in PVDF membrane. Conclusion：This work lay the foundation for further study on the function of iASPP.

关 键 词：p53凋亡刺激蛋白家族抑制成员 多克隆抗体 免疫原 免疫印迹 

分 类 号：R392-33[医药卫生—免疫学]