Establishment of anti-rat-Cu,Zn-superoxide dismutase monoclonal antibodies applied to a highly sensitive immunoassay and immunohistochemistry system.
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.
Nephrology and Rheumatology Division of Internal Medicine, Aichi Medical University, Yazako-Karimata, Nagakute, Japan. harutaka@amugw.aichi-med-u.ac.jp
Harutaka
H
Yamada
Yasukazu
Y
Yamada
Sachiko
S
Misao
Wataru
W
Kitagawa
Takafumi
T
Nomura
Takanari
T
Aoki
Naoto
N
Miura
Masato
M
Sakuma
Kazuhiro
K
Nishikawa
Arao
A
Futenma
Hirokazu
H
Imai
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D005260:Female; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D051381:Rats; D012680:Sensitivity and Specificity; D013482:Superoxide Dismutase / Q000276:immunology*
D000911:Antibodies, Monoclonal; D013482:Superoxide Dismutase