人血管能抑素免疫兔抗血清制备及其特性测定  

Preparation and Characteristic Test of the Rabbit Antiserum Against Human Canstatin

作　　者：贺国安[1] 罗进贤[1] 李瑞芳[1] 张添元[1] 吴群悦[1] 

机构地区：[1]中山大学基因工程教育部重点实验室,广东广州510275

出　　处：《中山大学学报（自然科学版）》2003年第6期134-136,共3页Acta Scientiarum Naturalium Universitatis Sunyatseni

基　　金：广东省自然科学基金资助项目(011207)

摘　　要：纯化的人血管能抑素包涵体进行SDS＿PAGE分离,切胶回收目的蛋白,与弗氏不完全佐剂充分乳化后,进行多点皮下注射免疫新西兰白兔,制备兔抗人血管能抑素抗血清,抗血清去除交叉反应抗体后,进行WesternBlotting分析抗血清与人血管能抑素及其N端和C端片段的抗原抗体反应特性。结果表明,经过4次加强免疫后,获得了高效价的兔抗人血管能抑素抗血清。抗血清去除交叉反应抗体后,WesternBlotting分析表明,它能与人血管能抑素发生特异的抗原抗体反应,同时还能与人血管能抑素的N端片段和C端片段发生特异性较弱的抗原抗体反应。证明制备的兔抗人血管能抑素抗血清能应用于人血管能抑素及其N端和C端片段免疫学研究中。The purified human canstatin was isolated by recovering the target band on the SDS-PAGE gel and used to immunize the New Zealand white rabbits by subcutaneous injection after emulsified with Freund's incomplete adjuvant. After removing its cross reacting antibodies, the antiserum against human canstatin was used to test its characteristic reaction with human canstatin and its N-terminal and C-terminal domains by Western blotting analyses. The results showed that high titer antiserum against human canstatin was obtained in immunized rabbits after four reinforced immunizations. The results of Western blotting analyses after removing the cross reacting antibodies showed that the rabbit antiserum against human canstatin specifically intensively reacted with human canstatin and faintly with its N-terminal and C-terminal domains. These results demonstrated the prepared rabbit antiserum against human canstatin can specifically react with human canstatin and its N-terminal and C-terminal domains, and can be applied in immunological stndy of N-terminal and C-terminal domains.

关 键 词：人血管能抑素 N端片段 C端片段 抗血清 兔 免疫 特性测定 

分 类 号：R392[医药卫生—免疫学] R338[医药卫生—基础医学]