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机构地区:[1]中南大学湘雅二医院血液科,长沙,410011,Zymed实验室,南三藩市,加利福尼亚94080,美国 [2]Zymed实验室,南三藩市,加利福尼亚94080,美国
出 处:《中南大学学报(医学版)》2005年第5期529-532,共4页Journal of Central South University :Medical Science
摘 要:目的:制备抗EphA4羧基端多肽的单克隆抗体,研究其免疫学特性。方法:体外合成EphA4羧基端多肽免疫小鼠,运用杂交瘤技术建立抗EphA4多肽单克隆抗体细胞株。通过免疫化学方法分析单抗的免疫学特性。结果:获得1株抗EphA4单克隆细胞,所分泌的抗体具有EphA4特异性,能识别人类、鸡与小鼠EphA4;可用于EphA4的ELISA、免疫沉淀、免疫印迹与免疫组织化学染色。结论:抗EphA4多肽单抗具备优良的免疫学特性,适用于多种免疫化学技术,是EphA4及其配体研究的重要工具。Objective To prepare monoclonal antibody (MAb) against carboxy-terminus of EphA4 and to analyze the immunological characteristics and significance of the antibody. Methods Mice were immunized with a chemically synthesized peptide that had been conjugated to KLH (keyhole limpet hemocyamin). A panel of antibodies specifically against EphA4 were obtained by hybridoma technique. The immunological properties and significance of the MAb were analyzed by immunological and immunochemistry techniques. Results A hybridoma cell line secreting anti-EphA4 MAb was established. The MAb reacted specifically with human, chicken and mouse EphA4, but did not cross-react with EphA5 and EphA7. The antibody could be used for immunochemical techniques such as ELISA, immunopreeipitation, Western blot and immunohistocnemistry. Conclusion The anti-EphA4 MAb generated by immunizing a synthetic peptide possesses excellent immunological properties. MAb can be applied for immunological and immunohistochemical purpose and will become an important tool in the study of EphA4 and its ligand.
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