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[Prokaryotic expression, purification and preparation of polyclonal antibody and immunohistochemistry analysis of
Yan Li1, Dong-dong Jiang, Cong Jin
1Department of Immunology Peking University School of Basic Medical Sciences, Beijing 100083, China.
Objective:
To express a thymocyte development related molecule RS21-C6 protein prepare its polyclonal antibodies and study its distribution in thymus by immunohistochemistry analysis.
Methods:
RS21-C6 recombinant protein was expressed in E.coli system and then purified by Ni-NTA chromatographic method. Polyclonal antibodies were prepared by immunized rabbits with the purified recombinant protein. Immunohistochemistry analysis was performed to show the distribution of RS21-C6 in mouse thymus tissue.
Results:
RS21-C6 recombinant protein was successfully expressed and purified. Then the high titer polyclonal antibody was prepared and its specificity was confirmed. The immunohistochemistry analysis demonstrated that RS21-C6 had an intense expression in the medullary region of thymus while scattered in the cortical region and the cortical medullary junction region of thymus. It also showed that RS21-C6 was co-expressed in thymocyte and thymic stromal cells.
Conclusion:
RS21-C6 molecule is broadly and highly expressed in thymus which suggests that this molecule may perform various functions in thymocytes at different developmental stages.
