title: Preparation and Characterization of a Polyclonal Antibody against Brominated Protein.
abstract: (Di)bromotyrosine is formed by the specific reaction of eosinophil peroxidase and can be used as an eosinophil activation marker. In the present study, an antibody for (di)bromotyrosine in proteins was prepared to investigate the pathogenesis of eosinophil-related diseases such as allergic responses. A rabbit polyclonal antibody was raised against brominated keyhole limpet hemocyanin. The specificity of the antiserum was investigated with an enzyme-linked immunosorbent assay (ELISA). The antiserum recognized brominated bovine serum albumin (BSA) and dibromotyrosine-conjugated BSA. The antiserum also reacted with chlorinated BSA and di-iodotyrosine-conjugated BSA. Moreover, the specificity of the antiserum was investigated using competitive ELISA. Dibromotyrosine and di-iodotyrosine inhibited the recognition of brominated BSA by the antiserum. However, the recognition of brominated BSA by the antiserum was not inhibited by bromotyrosine, chlorotyrosine, iodotyrosine, nitrotyrosine, aminotyrosine, phosphotyrosine, or tyrosine. These results suggested that the epitope of the antiserum is dihalogenated tyrosine. Immunohistochemically, the antiserum stained brominated rat eosinophils but not chlorinated or nitrated eosinophils. In conclusion, an antiserum for dihalogenated protein was prepared. It is expected that the antiserum will be useful for the analysis of the pathogenesis of allergic diseases such as asthma and atopic dermatitis.
authors.affiliation: Department of Environmental and Preventive Medicine, Graduate School of Medical Science, Kanazawa University, 13-1 Takara-machi, Kanazawa 920-8640, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Kambayashi
authors.forename: Keiki
authors.initials: K
authors.lastname: Ogino
authors.forename: Kei
authors.initials: K
authors.lastname: Takemoto
authors.forename: Takashi
authors.initials: T
authors.lastname: Imagama
authors.forename: Tomoko
authors.initials: T
authors.lastname: Takigawa
authors.forename: Shingo
authors.initials: S
authors.lastname: Kimura
authors.forename: Yuri
authors.initials: Y
authors.lastname: Hibino
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Hitomi
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Nakamura