In:
Bioscience, Biotechnology, and Biochemistry, Informa UK Limited, Vol. 81, No. 8 ( 2017-08-03), p. 1489-1496
Abstract:
Recombination activating gene 2 (RAG2) is necessary for immature B cell differentiation. Antibodies to human and rabbit RAG2 are currently commercially available, but antibodies to swine RAG remain unavailable to date. In this study, the swine RAG2 genes sequence was synthesized and then cloned into a pET-28a vector. The recombinant fusion protein was successfully expressed in E. coli, purified through nickel column chromatography, and further digested with Tobacco Etch Virus protease. The cleaved protein was purified by molecular-exclusion chromatography and named pRAG2. We used pRAG2 to immunize rabbits, collected the serum and purified rabbit anti-pRAG2 polyclonal antibodies. The rabbit anti-pRAG2 polyclonal antibodies were tested via immunofluorescence on eukaryotic cells overexpressing pRAG2 and also able to recognize pig natural RAG2 and human RAG2 protein in western blotting. These results indicated that the prepared rabbit anti-pRAG2 polyclonal antibodies may serve as a tool to detect immature B cell differentiation of swine.
Type of Medium:
Online Resource
ISSN:
0916-8451
,
1347-6947
DOI:
10.1080/09168451.2017.1340086
Language:
English
Publisher:
Informa UK Limited
Publication Date:
2017
detail.hit.zdb_id:
2110940-0
SSG:
12
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