新型半胱氨酰白三烯受体GPR17的多克隆抗体制备及其鉴定  

Preparation and identification of a polyclonal antibody against novel cysteinyl leukotriene receptor GPR17

作　　者：戚玲玲[1] 卢韵碧[1] 史文珍[1] 赵春贞[1] 张咏梅[1] 陈丽萍[2] 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]衢州学院医学院药理学教研室,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4期357-361,共5页Journal of Zhejiang University(Medical Sciences)

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

摘　　要：目的:制备新型半胱氨酰白三烯受体GPR17的多克隆抗体(pAb),并鉴定其免疫学特性。方法:以KLH偶联的GPR17多肽免疫家兔制备其pAb,用间接ELISA法测定抗体效价,阻断ELISA测定抗体敏感性及特异性,同时以新制备的抗体做Western blot,检测GPR17的组织表达。结果:通过免疫家兔获得的抗GPR17 pAb,ELISA测定显示其抗体效价最高可达到1/16 364,并对CysLT1受体和CysLT2受体基本无交叉反应,具有较好的特异性。Western blot结果显示,GPR17在大鼠脑和心脏组织中有较高的表达,其分子量在43 kD左右。结论:制备的GPR17 pAb具有较高的效价和较好的特异性,能满足Western blot检测GPR17的要求。Objective： To prepare and identify a polyclonal antibody （pAb） against GPR17,a novel cysteinyl leukotriene receptor. Methods： Rabbits were immunized with KLH-coupled GPR17 peptide to prepare the pAb. The titer of the pAb in rabbit plasma was detected by indirect ELISA,and the specificity of the pAb was tested by antigen blockade. GPR17 tissue distribution was detected by Western blot with the pAb. Results： The pAb showed a titer as high as 1 ： 16 364,and was not cross-reacted with the antigens of CysLT1 and CysLT2 receptors. A higher expression of GPR17 in the rat brain and heart was detected using the newly prepared pAb. The molecular weigh of GPR17 protein was about 43 kD. Conclusion： The prepared GPR17 pAb has high sensitivity and specificity,and can be used in Western blot for detecting GPR17.

关 键 词：受体 白三烯 半胱氨酸 GPR17 半胱氨酰白三烯受体 多克隆抗体 

分 类 号：S852.43[农业科学—基础兽医学]