[Prokaryotic expression of the extracellular region of human CD1d and preparation of its polyclonal antibody].
OBJECTIVE
To explore the prokaryotic expression of the extracellular region of human CD1d (hCD1d) and prepare its polyclonal antibody.


METHODS
The gene encoding the extracellular region of hCD1d was amplified by PCR and cloned into prokaryotic expression vector pET28, then expressed in E.coli BL21 (DE3) with IPTG induction. The recombinant protein was purified by Ni2+-NTA agarose column and then used as immunogen to immunize the mouse. The generated polyclonal antibody was evaluated by ELISA, Western blot and immunohistochemical staining (IHC), respectively.


RESULTS
The recombinant extracellular region of hCD1d was successfully expressed and purified. The polyclonal antibody with high titer and high specificity was obtained, which could recognize the native hCD1d in the human small intestinal tissues.


CONCLUSIONS
The recombinant extracellular region of hCD1d has been obtained. The antibody with high titer and high specificity against the extracellular region of hCD1d from the mouse has been successfully prepared, which lays a foundation for further research into the detection and functional study of CD1d.
Chen
Zhang-quan
ZQ
Department of Clinical Immunology, Guangdong Medical College, Zhanjiang 524023, China. czqhk@126.com
Huang
Zhen
Z
Liang
Xiao-dong
XD
Lu
Tian-tian
TT
He
Tian-wen
TW
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D055610:Antigens, CD1d / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D016133:Polymerase Chain Reaction
D000906:Antibodies; D055610:Antigens, CD1d; C526877:CD1D protein, human