小鼠PC-1蛋白兔多克隆抗体的制备及初步应用  

Generation and Preliminary Application of Polyclonal Antibody Against Murine PC-1 Protein

作　　者：万里川[1] 周建光[1] 李杰之[1] 孙玉龙[1] 刘春丽[2] 

机构地区：[1]军事医学科学院生物工程研究所 [2]军事医学科学院毒物药物研究所,北京100850

出　　处：《中国生物工程杂志》2003年第12期95-98,共4页China Biotechnology

基　　金：国家自然科学基金资助项目 ( 3 0 0 70 2 96)

摘　　要：目的 :获得小鼠PC 1蛋白的兔多克隆抗体 ,检测其在小鼠器官中的表达谱。方法 :以纯化后的小鼠PC 1蛋白及其N端 45个氨基酸与GST的融合蛋白 (GST MPC 1、GST MPC 1 45 )为抗原 ,制备了它们的兔多克隆抗体。进一步应用饱和硫酸铵沉淀法、DEAE5 2和ProteinA亲和层析法对这两个抗体进行了纯化 ,Westernblot检测了PC 1蛋白在昆明白小鼠七个组织中的表达情况。结果 :两个兔多克隆血清的效价分别为 1∶2 0 0 0 0和 1∶2 0 0 0 0 0 ,并能与人工合成的人PC 1蛋白N端的 46个氨基酸发生良好的交叉反应。PC 1蛋白在结肠中有表达 ,在肾中有微弱表达 ,而在心、肺、肝、胃、脾中没有表达。结论 :获得小鼠PC 1蛋白的兔多克隆抗体 ,PC 1蛋白在结肠中有表达 ,在肾中有微弱表达 ,而在心、肺、肝、胃。Objective: To produce rabbit polyclonal antibody of murine PC 1 protein and detect its expression profile in mouse organs.Methods: Rabbit polyclonal antibodies were produced by utilizing GST fusion proteins of murine PC 1 protein and its N terminal 45 amino acids(GST MPC 1 and GST MPC 1 45),which were purified by Glutamine Sepharose 4B,as antigens.The antisera were purified by precipitation of saturated ammonium sulfate,DEAE52,and Protein A affinity chromatography.Western blot was used to examine murine PC 1 protein expression spectrum.Results:The two rabbit antisera had a title of 1∶20000 and 1∶200000,respectively,and had good cross reactions with manmade human PC 1 protein N terminal 46 amino acids.The expression of murine PC 1 protein has been detected in colon and at low level in kidney,but not in heart,lung,liver,gastric gland and spleen.Conclusion:Rabbit polyclonal antibody has been generated and the expression of murine PC 1 protein has been detected in colon and at low level in kidney,but not in heart,lung,liver,gastric gland and spleen.

关 键 词：多克隆抗体 前列腺癌 PC-1基因 免疫 

分 类 号：R737.25[医药卫生—肿瘤]