Production of polyclonal antibody against interleukin-33 and assessment of its distribution in murine liver and lung.
Interleukin (IL)-33 is the latest member of IL-1 cytokine family. In this study, the cloning, expression, purification, and polyclonal antibody preparation of mouse IL-33 were described. The coding region of IL-33 mature protein was cloned into the prokaryotic expression vector pET-44. The recombinant protein, IL-33 containing a hexahistidine tag in the C-terminal, was expressed in Escherichia coli. The expressed soluble protein was purified by immobilized metal-ion affinity chromatography using Ni2+-nitrilotriacetic acid agarose. The rabbits were immunized with the purified recombinant protein. The obtained antiserum was precipitated by saturated ammonium sulfate and then purified by Protein A affinity chromatography. The sensitivity and specificity of the antibodies were evaluated by enzyme-linked immunosorbent assay and immunohistochemistry. The high titer (1 : 32000) polyclonal antibodies with high specificity were obtained by immunizing rabbits with the purified recombinant protein. Significant expression of IL-33 was seen in mouse liver and lung tissues determined with the anti-IL-33. The production of the polyclonal antibody against IL-33 provides a good tool for studying the biofunctions of IL-33.
Liu
Xiaojin
X
Institute of Inflammation and Immune Diseases, Shantou University Medical College, Shantou 515041, China.
Wu
Yan
Y
Li
Mingcai
M
Chen
Sui
S
Zhou
Yanchun
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology*; D002478:Cells, Cultured; D000067596:Interleukin-33; D007378:Interleukins / Q000276:immunology*; D008099:Liver / Q000276:immunology*; D008168:Lung / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D015202:Protein Engineering / Q000379:methods*; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; C506535:Il33 protein, mouse; D000067596:Interleukin-33; D007378:Interleukins; D011994:Recombinant Proteins