Preparation of Rabbit Polyclonal Antibody against ELF3 and Its Immunolocalization in Mice
10.3870/j.issn.1672-0741.2010.01.002
- VernacularTitle:ELF3多克隆抗体的制备及其免疫定位
- Author:
Zhijun WANG
;
Ying CHANG
;
Wei WU
;
Jinjian YAO
;
Xianxian HE
;
Jusheng LIN
;
Xuhu SONG
- Publication Type:Journal Article
- Keywords:
embryonic liver fodrin 3;
antibody preparation;
immunolocalization
- From:
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
2010;39(1):6-8,17
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare and identify rabbit polyclonal antibody against embryonic liver fordrin 3(ELF3),and investigate the distribution of ELF3 in mice tissue.Methods ELF3 specific N-terminal peptide was synthesized,and conjugated to Keyhole limpet hemocyanin(KLH)as immunogen.The ELF3-KLH complex was injected into rabbits subcutaneously,and then ELF3 antibody was purified using affinity chromatography.The titer of the antibody was evaluated by ELISA.The specificity of antibody against ELF3 and immunolocalization of ELF3 were evaluated by using Western blot and immunohistochemistry.Results Rabbit polyclonal antibody against ELF3 was prepared by the immunization of ELF3-KLH complex.ELISA and Western blot results showed the antibody against ELF3 had high titer and specificity.Western blot and immunohistochemical studies demonstrated ELF3 was expressed in the mouse heart,liver,brain and kidney tissue,particular on the cell membrane.Conclusion The preparation of polyclonal antibody against ELF3 was successful due to its high titer and specificity;ELF3 was expressed in the mice heart,liver,and kidney,particular on the cell membrane.It will provide an excellent tool for further study on the ELF3 function.
