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.
| Original language | English |
|---|---|
| Pages (from-to) | 95-103 |
| Number of pages | 9 |
| Journal | Journal of Clinical Biochemistry and Nutrition |
| Volume | 44 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - Jan 2009 |
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Keywords
- (Di)bromotyrosine
- Allergic disease
- Eosinophil activation marker
- Oxidative stress
- Polyclonal antibody
ASJC Scopus subject areas
- Clinical Biochemistry
- Medicine (miscellaneous)
- Nutrition and Dietetics
Cite this
Preparation and characterization of a polyclonal antibody against brominated protein. / Kambayashi, Yasuhiro; Ogino, Keiki; Takemoto, Kei; Imagama, Takashi; Takigawa, Tomoko; Kimura, Shingo; Hibino, Yuri; Hitomi, Yoshiaki; Nakamura, Hiroyuki.
In: Journal of Clinical Biochemistry and Nutrition, Vol. 44, No. 1, 01.2009, p. 95-103.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Preparation and characterization of a polyclonal antibody against brominated protein
AU - Kambayashi, Yasuhiro
AU - Ogino, Keiki
AU - Takemoto, Kei
AU - Imagama, Takashi
AU - Takigawa, Tomoko
AU - Kimura, Shingo
AU - Hibino, Yuri
AU - Hitomi, Yoshiaki
AU - Nakamura, Hiroyuki
PY - 2009/1
Y1 - 2009/1
N2 - (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.
AB - (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.
KW - (Di)bromotyrosine
KW - Allergic disease
KW - Eosinophil activation marker
KW - Oxidative stress
KW - Polyclonal antibody
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UR - http://www.scopus.com/inward/citedby.url?scp=60849115833&partnerID=8YFLogxK
U2 - 10.3164/jcbn.08-196
DO - 10.3164/jcbn.08-196
M3 - Article
VL - 44
SP - 95
EP - 103
JO - Journal of Clinical Biochemistry and Nutrition
JF - Journal of Clinical Biochemistry and Nutrition
SN - 0912-0009
IS - 1
ER -