Production of Polyclonal Antibody against Interleukin-33 and Assessment of Its Distribution in Murine Liver and Lung

الملخص EN

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.