Objective:To prepare and identify the monoclonal antibody(mAb) against protease activated receptor-2(PAR-2). Methods:BLAB/c mice were immunized with specific PAR-2 polypeptide by multiple-site hypodermic injection. The spleen cells of immunized mice were fused with NS-1 myeloma cells. Indirect ELISA was used to screen the hybridoma culture supernants. The specificity of mAb were characterized by ELISA,Dot blot,immunohistochemistry,flow cytometry and confocal laser scanning microscopy(CLSM). Results:One hybridoma cell line against PAR-2 was successfully obtained, this mAb was IgM type and the titer of antibody in supernants was 1 ∶ 16. Dot blot analysis showed that the mAb could specifically bind to PAR-2. Immunohistochemical staining revealed that the reactivity of mAb to epithelial cells and smooth muscle cells in lung, epithelial cells and lymphocyte-like cells in colon, lymphocytes in tonsil and squamous epithelial cells in skin tissue. The result of flow cytometry showed that the mAb recognized PAR-2 in A549 cells. CLSM examination confirmed that the fluorescent markers were localized in both cytomembrane and cytoplasm of A549 cells. Conclusion:The mAb against PAR-2 is a useful tool for investigate PAR-2 related disease