Cloning and purification of paired immunoglobin-like type 2 receptor beta and preparation of its polyclonal antibody
- Author:
Jian-Zhong LI
1
Author Information
1. Department of Biochemical Pharmacy
- Publication Type:Journal Article
- From:
Academic Journal of Second Military Medical University
2006;27(9):1011-1013
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To express and purify paired immunoglobin-like type 2 receptor beta (PILRβ) and prepare the polyclonal antibody against its protein. Methods: The PILRβ 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 PILRβ gene was then expressed in E. coli by IPTG induction and the product was purified by Ni2+-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 PILRβ protein (Mr 42 000) was obtained. The prepared polyclonal antibody was highly specific and had a titer of 1:10 000. Conclusion: PILRβ gene is successfully cloned and the purified PILRβ protein is expressed. Polyclonal antibody against PILRβ(with high titer and specificity) is successfully obtained, which provides a basis for further study on PILRβ.
