摘要
目的:制备兔抗人脱嘌呤/脱嘧啶核酸内切酶多克隆抗体并进行鉴定。方法:以纯化的全长APE1蛋白为抗原,采用改良的快速免疫法制备兔抗人APE1多克隆抗体并进行特异性亲和纯化,ELISA、Western blot和免疫组化检测制备抗体的效价和特异性。结果:ELISA检测显示该抗体的效价达到1∶128000,相对亲和力常数为8.96×10-6mol/L。Western blot显示该抗体能与APE1蛋白特异性结合,免疫组化检测显示阳性反应产物主要定位于细胞核。此外,该抗体还可用于小鼠和大鼠组织APE1蛋白的检测。结论:制备的多克隆抗体特异性和效价良好,不仅为今后深入研究人APE1蛋白的性质与功能提供了有用的实验工具,还可用于检测小鼠和大鼠组织中APE1蛋白的表达。
AIM： To prepare a rabbit anti-human apurinic/apyrimidinic endonuclease polyclonal antibody and then have its characterization analyzed, METHODS： A rabbit polyclonal antibody was prepared through a modified rapid immune procedure and then it was purified using a specific avidity column. The efficacy of this antibody was detected by ELISA, Western blot and immunohistochemistry. RESULTS ： The titer of the antiserum determined by ELISA was up to 1：128 000. The kaff value of the antibody was 8.96 × 10^-6 mol/L. Western blot analysis showed the antibody was of high specificity. The final purified antibody was highly specific to native form of APE1 protein in mice and rats. CONCLUSION： The rabbit anti-human APE1 polyclonal antibody with high titer and specificity has been successfully prepared. It can be used not only to elucidate the roles of APE1 protein in many important cellular procedures, but also to detect the expression of the APE1 protein in mice and rats.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2009年第4期325-327,331,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金资助面上项目(30872975)
