应用Protein A亲和层析法制备及纯化R1 JHL单克隆抗体  

Preparation and purification of mouse monoclonal antibody against NMDAR1 by ProteinA-Sepharose CL-4B affinity chromatography

作　　者：李桂新[1] 孙长凯[2] 程桂芝[3] 罗建红[4] 李芳[3] 徐淑君[4] 

机构地区：[1]大连医科大学附属第一医院检验科,辽宁大连116011 [2]大连医科大学脑疾病研究所,辽宁大连116044 [3]大连医科大学免疫学教研室,辽宁大连116044 [4]浙江大学医学院分子中心,浙江杭州310031

出　　处：《大连医科大学学报》2008年第2期188-190,共3页Journal of Dalian Medical University

摘　　要：[目的]制备、纯化N-甲基D-门冬氨酸受体(NMDAR,NR)主亚基RIJHL单克隆抗体。[方法]NR1杂交瘤细胞腹腔注射BalB/C小鼠制备抗体,Protein A-Sepharose CL-4B亲和层析纯化单抗,Western-blot鉴定该单抗特异性。[结果]纯化后单抗识别大鼠脑组织膜蛋白中约115 kD大小的单一条带。[结论]制备、纯化了具有高度特异性的抗NR1单克隆抗体,为进一步研究NMDA受体提供了工具。[Objective] To prepare and purify the monoclonal antibody against NMDAR1, [Methods] NR1 Hybridoma cells were injected into the abdominal cavity of BaIB/C mice to prepare the monoclonal antibody against NMDAR1, The antibodies were purified by ProteinA - Sepharose CL-4B affinity chromatography and its specificity were evaluted by Western blot, [Results] Western blot comfired that the antibodies specifically recognized the NR1 membrane protein extracted from rat brain tissues. [Conclusion] The purified antibodies can be employed to the further study of NMDAR.

关 键 词：NMDA受体 杂交瘤细胞 单克隆抗体制备纯化 WESTERN-BLOT 

分 类 号：R392[医药卫生—免疫学]