半胱氨酰白三烯受体2多克隆抗体制备及其鉴定  

Preparation and identification of polyclonal antibody against cysteinyl leukotriene receptor 2

作　　者：陈丽萍[1,2] 赵春贞[1] 史文珍[1] 戚玲玲[1] 卢韵碧[1] 张咏梅[1] 张丽慧[3] 方三华[1] 鲍建芳[4] 沈建根[4] 魏尔清[1] 

机构地区：[1]浙江大学医学院药理学系,浙江杭州310058 [2]衢州职业技术学院医学系机能教研室,浙江衢州324000 [3]杭州师范大学基础医学院药理学教研室,浙江杭州310036 [4]浙江大学医学院免疫研究所,浙江杭州310058

出　　处：《浙江大学学报（医学版）》2009年第6期591-597,共7页Journal of Zhejiang University(Medical Sciences)

基　　金：国家自然科学基金资助项目(30772561);浙江省自然科学基金资助项目(Y204210)

摘　　要：目的:制备半胱氨酰白三烯受体2(CysLT2)多克隆抗体,并鉴定其免疫学特性及应用。方法:以KLH偶联的CysLT2受体多肽免疫家兔制备抗体,用间接ELISA法测定抗体效价,以抗原阻断法测定抗体敏感性及特异性;同时,以新制备的抗体进行Western blot和免疫组化,检测CysLT2受体的组织表达。结果:ELISA测定显示,获得的家兔抗CysLT2受体抗体的效价大于1/1 047 296,对CysLT2受体的特异性强,对CysLT1受体和GPR17基本无交叉反应。Western blot结果显示,CysLT2受体在大鼠、小鼠肾、脑和肺中有较高的表达,其分子量在40 kD左右。免疫组化结果表明,CysLT2受体主要表达在大鼠神经元,也表达在部分星形胶质细胞。结论:制备的CysLT2受体多克隆抗体具有高效价和高特异性,能满足Western blot和免疫组化检测的要求。Objective： To prepare and identify a polyclonal antibody against cysteinyl leukotriene receptor （CysLT2 receptor）. Methods： Rabbits were immunized with KLH-eoupled CysLT2 receptor peptide to prepare the polyclonal plasma was detected by indirect ELISA,and the antibody （pAb）. The titer of the pAb in rabbit specificity of the pAb was tested by antigen blockade. The tissue distribution of CysLT2 receptor was detected by Western blot and immunohistochemistry with the prepared pAb. Results： The pAb showed a titer higher than 1/ 1047296,and was specific to CysLT2 receptor,without cross-reaction with the antigens of CysLT1 receptor and GPR17. A higher expression of CysLT2 receptor in kidney,brain and lung of rats and mice was detected by Western blot analysis using the prepared pAb. The molecular weight of CysLT2 receptor protein was about 40 kD. Immunohistochemical examination showed that CysLT2 receptor was expressed mainly in the neuron, and partly in astrocytes in rat brain. Conclusions： The prepared CysLT2 receptor pAb has high sensitivity and specificity,and can be used in Western blot and immunohistochemistry.

关 键 词：半胱氨酸/遗传 受体 白三烯/遗传 半胱氨酰白三烯受体 CysLT2受体 多克隆抗体 

分 类 号：Q78[生物学—分子生物学]