[Prokaryotic expression, purification and preparation of polyclonal antibody and immunohistochemistry analysis of RS21-C6 molecule].
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.


CONCLUSIONS
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.
Li
Yan
Y
Department of Immunology  Peking University School of Basic Medical Sciences, Beijing 100083, China.
Jiang
Dong-dong
DD
Jin
Cong
C
Liu
Yuan-feng
YF
Chen
Wei-feng
WF
D000818:Animals; D015153:Blotting, Western; D007150:Immunohistochemistry; D051379:Mice; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000302:isolation & purification* / Q000494:pharmacology; D013601:T-Lymphocytes / Q000502:physiology*; D013950:Thymus Gland / Q000737:chemistry / Q000254:growth & development*
D011993:Recombinant Fusion Proteins