兔抗鼠CaV1.2羧基末端多克隆抗体的制备和鉴定  

Generation and characterization of rabbit polyclonal antiserum against C terminus of rat CaV1.2

作　　者：许多[1] 刘大力[2] 赵守亮[1] 粱景平 

机构地区：[1]同济大学附属口腔医院,上海200072 [2]上海交通大学附属口腔医院,上海200011

出　　处：《牙体牙髓牙周病学杂志》2011年第4期229-232,213,共5页Chinese Journal of Conservative Dentistry

基　　金：国家自然科学基金资助项目(81070825);上海市科委医学引导项目(10411964600)

摘　　要：目的:制备高效价的兔抗鼠钙通道CaV1.2羧基末端多克隆抗血清。方法:人工合成鼠L型钙离子通道CaV1.2羧基末端短肽;用该短肽与马来酰胺活化匙孔血蓝蛋白(Keyhole limpet hemocyanin,KLH)偶联免疫兔获得抗血清;利用饱和硫酸铵盐析和亲和层析法获得纯化后抗血清;酶联免疫吸附法实验测定抗体效价;斑点杂交实验测定抗血清特异性。结果:经免疫得到兔抗鼠L型钙离子通道CaV1.2羧基末端多克隆抗血清;ELISA和斑点杂交实验证实兔抗鼠CaV1.2羧基末端多克隆抗体可以特异性识别CaV1.2羧基末端多肽。结论:本实验成功制备纯化抗CaV1.2羧基末端多克隆抗体并且可以用于后续CaV1.2羧基末端水解片段的功能研究。AIM：To obtain rabbit anti-rat C terminus of calcium channel CaV1.2 polyclonal antiserum. MEHODS：The short peptides from C terminus of rat CaV1.2 were synthesized,which were used to cross-link with Keyhole limpet hemocyanin,KLH.New Zealand white rabbits were immunized by synthesized peptides-KLH to obtain polyclonal antiserum.The antibody was purified by ammonium sulfate and affinity chromatography and characterized by ELISA and dot blot.RESULTS：Antiserum against C terminus of rat CaV1.2 was obtained in immunized rabbits. ELISA and dot blot identified that the antiserum could recognize the peptides of C terminus of rat CaV1.2.CONCLUSION： The polyclonal antiserum has been generated successfully,which can be used to explore the potential function of the C terminus of L-type calcium channel.

关 键 词：钙离子通道 抗体制备 酶联免疫吸附法 蛋白斑点杂交 

分 类 号：R780.2[医药卫生—口腔医学]