兔抗鼠CaV1.2羧基末端多克隆抗体的制备和鉴定

摘要

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 CaVl.2 was obtained in immunized rabbits.ELISA and dot blot idertified that the antiserum could recognize the peptides of C terminus of rat CaVI.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.%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.