抗微小膜泡蛋白16多克隆抗体的制备、纯化及其在亚细胞定位中的应用
[Preparation and purification of polyclonal antibody against microneme protein 16 and its application in subcellular localization].
作者信息
Dong-Dong Wei, Long-Jiang Wang, Jin Li, Yong Cui, Kun Yin, Bing-Cheng Huang, Qing-Kuan Wei, Zhan Lei, Hui Sun
机构信息
Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Diseases, Jining 272033, China.
Co-first author.
出版信息
Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi. 2018 Aug 16;30(4):440-442. doi: 10.16250/j.32.1374.2017216.
OBJECTIVE
To prepare and purify the rabbit anti-TgMIC16 polyclonal antibody, so as to apply it in subcellular localization.
METHODS
New Zealand white rabbits were immunized with purified recombinant TgMIC16 mixing with the same volume of Freund's adjuvant for three times, respectively. The rabbit serum was collected on the 14th day after the last immunization. The polyclonal antibody in rabbit serum was purified with Protein A affinity purification column. ELISA and Western blotting were used to detect the antibody titer and specificity of polyclonal antibody. The polyclonal antibody was used to the localization of TgMIC16 by the immunofluorescence method.
RESULTS
Indirect ELISA showed that the antibody titer was 1∶512 000. Western blotting showed that the recombinant TgMIC16 protein was recognized by the specific polyclonal antibody. IFA showed that TgMIC16 was located in the microneme of .
CONCLUSIONS
The rabbit anti-TgMIC16 is prepared and purified, and successfully applied to immunofluorescence localization of TgMIC16 in .
目的
制备并纯化兔抗TgMIC16多克隆抗体，以便将其应用于亚细胞定位。
方法
用纯化的重组TgMIC16与等体积弗氏佐剂混合，分别对新西兰白兔进行3次免疫。末次免疫后第14天采集兔血清。用蛋白A亲和纯化柱纯化兔血清中的多克隆抗体。采用ELISA和Western印迹法检测多克隆抗体的效价和特异性。采用免疫荧光法用多克隆抗体对TgMIC16进行定位。
结果
间接ELISA显示抗体效价为1∶512 000。Western印迹显示重组TgMIC16蛋白能被特异性多克隆抗体识别。免疫荧光分析显示TgMIC16位于[此处原文缺失具体位置信息]的微线体中。
结论
制备并纯化了兔抗TgMIC16多克隆抗体，并成功将其应用于[此处原文缺失具体位置信息]中TgMIC16的免疫荧光定位。
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