Preparation and Characterization of Polyclonal Antibody Against Xenopus PAPC
- VernacularTitle:非洲爪蟾PAPC多克隆抗体的制备及其特异性鉴定
- Author:
Ruiying HU
;
Peng XU
;
Yuelei CHEN
;
Xin LOU
;
Xiaoyan DING
- Publication Type:Journal Article
- Keywords:
Xenopus laevis;
Paraxial Protocadherin (PAPC);
prokaryotic expression;
polyclonal antibody;
Western blotting;
immunofluorescent staining
- From:
Progress in Biochemistry and Biophysics
2007;34(2):222-228
- CountryChina
- Language:Chinese
-
Abstract:
Xenopus Paraxial Protocadherin (PAPC), which was initially identified in a screen for genes present in the Spemann organizer of Xenopus embryos, is required for gastrulation, somitogenesis and otic vesicle formation. In order to investigate its function in various developmental events, an antibody was prepared which could specifically recognize Xenopus PAPC. Glutathione S transferase (GST) expression system was used to express the fusion protein GST-PAPC. Rabbits were immunized with GST-PAPC Western blotting analysis of FL-PAPC transfected HEK 293T cells lysates, which could be specifically blocked by pre-adsorption of prokaryotic expressed GST-PAPC fusion protein. Furthermore, by using immunofluorescence analysis the polyclonal antibody recognized membrane-bound PAPC in FL-PAPC transfected 293T cells and Xenopus animal cap cells. By Western blotting analysis,the endogenous 150 ku PAPC protein was detected in Xenopus embryos using the anti-PAPC antibody. Take together it could be concluded that a polyclonal antibody specifically against Xenopus PAPC was developed.
