Preparation and purification of polyclonal antibody against Toxoplasma gondii microneme protein 16 and its application in subcellular localization
10.16250/j.32.1374.2017216
- VernacularTitle:弓形虫TgMIC16多克隆抗体制备纯化及其在亚细胞定位中的应用
- Author:
Dong-Dong WEI
1
;
Long-Jiang WANG
2
;
Jin LI
1
;
Yong CUI
1
;
Kun YIN
1
;
Bing-Cheng HUANG
1
;
Qing-Kuan WEI
1
;
Zhan LEI
3
;
Hui SUN
1
Author Information
1. Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Diseases, Jining 272033, China
2. Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Diseases, Jining 272033, China; Co-first author
3. Shandong Academy of Medical Sciences, Shandong Laboratory Animal Center, China
- Publication Type:Journal Article
- Keywords:
Toxoplasma gondii;
Microneme protein 16;
Polyclonal antibody;
Immunofluorescence localization
- From:
Chinese Journal of Schistosomiasis Control
2018;30(4):440-442
- CountryChina
- Language:Chinese
-
Abstract:
To prepare and purify the rabbit anti-TgMIC16 polyclonal antibody, so as to apply it in subcellular localization.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.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 Toxoplasma gondii.The rabbit anti-TgMIC16 is prepared and purified, and successfully applied to immunofluorescence localization of TgMIC16 in T. gondii.
