Eukaryotic expression and preparation of polyclonal antibody for osteoclast inhibitory lectin related protein
10.3969/j.issn.1000-484X.2014.10.016
- VernacularTitle:破骨细胞抑制凝集素相关蛋白2的真核表达及多克隆抗体制备
- Author:
Suxia WU
;
Lihui CHAI
;
Zhanzheng WANG
;
Guangchao LIU
;
Wenzhi TIAN
;
Yuanfang MA
- Publication Type:Journal Article
- Keywords:
C-type lectin-related protein;
Eukaryotic expression;
Polyclonal antibody;
Dendritic cell
- From:
Chinese Journal of Immunology
2014;(10):1374-1377,1392
- CountryChina
- Language:Chinese
-
Abstract:
Objective:In order to get recombinant protein OCILRP 2-Fc and anti-OCILRP2 antibody for further study of OCILRP2.Methods:Eukaryotic expression vector pIg-CD5-OCILRP2 which fused with extracellular domain of OCILRP 2 and human IgG1 Fc fragment was constructed.G418 was used for stable expression cell strain after pIg-CD5-OCILRP2 transfected into CHO cells.Recombinant protein OCILRP 2-Fc purified from CHO cell supernatant was used to immunize rabbit and anti-OCILRP2 polyclonal antibody was purified from rabbit serum by using protein G column.Results: ELISA data showed that we got a high-titer anti-serum and anti-OCILRP2 antibody purified from the rabbit serum.Western blot indicated this antibody could specifically bind to OCILRP 2-Fc and OCILRP2 in NIH/3T3 lysate.OCILRP2 expression in murine bone marrow derived dendritic cells ( DC) was detected by this polyclonal antibody ,too.Compared to immature DC ,OCILRP2 expression was elevated in LPS induced-mature DC.Conclusion: This study has offered an available tool and provided a clue for further study of the roles of OCILRP 2 in immune response.
