| 兔抗人DR5分子多克隆抗体的制备与初步应用 |
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| 引用本文： | 白慧玲,裴景堂,杜耀武,李淑莲,刘广超,马远方.兔抗人DR5分子多克隆抗体的制备与初步应用[J].免疫学杂志,2008,24(6):634-637. |
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| 作者姓名： | 白慧玲 裴景堂 杜耀武 李淑莲 刘广超 马远方 |
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| 作者单位： | 河南大学免疫学研究所河南省天然药物与免疫工程重点实验室;河南大学附属淮河医院; |
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| 基金项目： | 河南省教育厅自然科学基础研究(200510475039);;国家自然科学基金项目(30571697) |
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| 摘 要： | 目的获得兔抗人DR5的多克隆抗体,并对多克隆抗体进行初步鉴定。方法皮下注射DR5免疫日本大耳白兔,加强免疫得到抗血清,应用蛋白G纯化获得多克隆抗体。对纯化的抗体进行ELISA、免疫印迹、细胞凋亡活性等初步鉴定。结果经纯化后的抗体效价为1∶32000;Western blot分析该抗体能够识别相对分子质量26000的sDR5;可以明显导致肿瘤细胞Jurkat凋亡,Western blot检测到随着抗体诱导时间的延长Jurkat细胞Bcl-2蛋白表达减少,Cyt-C、Bax、有活性的caspase3和caspase9蛋白的表达增加。结论所制备的多克隆抗体可应用于ELISA、免疫印迹和诱导肿瘤细胞凋亡活性及机制等实验,为确定DR5分子的组织及细胞分布和定位等提供了有力的工具。
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| 关 键 词： | DR5 多克隆抗体 凋亡 |
Generation and preliminary application of rabbit antihuman polyclonal antibodies against DR5 |
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| BAI Hui-ling,PEI Jing-tang,DU Yao-wu,LI Shu-lian,LIU Guang-chao,MA Yuan-fang.Generation and preliminary application of rabbit antihuman polyclonal antibodies against DR5[J].Immunological Journal,2008,24(6):634-637. |
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| Authors: | BAI Hui-ling PEI Jing-tang DU Yao-wu LI Shu-lian LIU Guang-chao MA Yuan-fang |
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| Institution: | Institute of Immunology;Henan University;Henan Kaifeng 475004;China |
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| Abstract: | Objective To generate and identify the rabbit polyclonal antibodies against sDR5. Methods Rabbits were immunized with sDR5 for 3 times. The polyclonal antibodies from the collected antiserum were purified with protein G. Specialty of the polyclonal antibodies was analyzed by indirect-ELISA; apoptotic effects of the polyclonal antibodies on Jurkat cells were detected by FCM analysis; Bcl 2,CytC,caspase3,Bax,and caspase9 were detected with Western blotting. Results The titers of purified polyclonal antibodies... |
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| Keywords: | DR5 |
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