Establishment of Anti-Rat–Cu,Zn-Superoxide Dismutase Monoclonal Antibodies Applied to a Highly Sensitive Immunoassay and Immunohistochemistry System
Harutaka Yamada, Yasukazu Yamada, Sachiko Misao, Wataru Kitagawa, Takafumi Nomura, Takanari Aoki, Naoto Miura, Masato Sakuma, Kazuhiro Nishikawa, Arao Futenma, Hirokazu Imai · Hybridoma and Hybridomics · 2004
The superoxide anion has been implicated in a wide range of diseases. The major protector against superoxide anion in the cell cytosol is Cu,Zn-superoxide dismutase (Cu,Zn-SOD). In this study, anti rat Cu,Zn-SOD was established in murine monoclonal antibodies for the first time. These antibodies were applied to both a highly sensitive EIA system in serum and immunohistochemical methods for detection in gastric mucosa tissues. The proposed EIA method had a high sensitivity within the assay range, 10-300 pg/mL, good percentage, 96.9 +/- 5.60%, and good reproducibility; within-day assay CV = 8.6-10.2%, between-day assay CV = 6.5-11.7%. Inmmunohistochemically, Cu,Zn-SOD localized in the esophagus epithelial cells, gastric oxyntic cells, surface of the gastric lumen side in the small intestine and colonic epithelial cells. The establishment of anti-rat CuZn-SOD monoclonal antibody allows both specific analysis of immunoquantitation in rat Cu,Zn-SOD and highly specific detection of Cu,Zn-SOD location by immunohistochemical methods.