Cloning and purification of paired immunoglobin-like type 2 receptor beta and preparation of its polyclonal antibody
- VernacularTitle:人Ⅱ型成对免疫球蛋白样受体?基因(PILR?)的克隆、表达、纯化及其多克隆抗体的制备
- Author:
Jianzhong LI
- Publication Type:Journal Article
- Keywords:
paired immunoglobin-like type 2 receptor ?;
cloning,molecular;
gene expression;
polyclonal antibody
- From:
Academic Journal of Second Military Medical University
2001;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To express and purify paired immunoglobin-like type 2 receptor beta(PILR?) and prepare the polyclonal antibody against its protein.Methods: The PILR? cDNA was amplified from the total RNAs extracted from human peripheral blood cells by RT-PCR and was cloned into pET-32a vector.The vector harboring PILR? gene was then expressed in E.coli by IPTG induction and the product was purified by Ni~(2+)-NTA affinity chromatography.Polyclonal antibody was prepared by immunizing rabbits with the purified protein.The titer and specificity of the antibody were examined by ELISA and Western blot,respectively.Results: Highly purified PILR? protein(Mr 42 000) was obtained.The prepared polyclonal antibody was highly specific and had a titer of 110 000.Conclusion: PILR? gene is successfully cloned and the purified PILR? protein is expressed.Polyclonal antibody against PILR?(with high titer and specificity) is successfully obtained,which provides a basis for further study on PILR?.
