Use of commercial antibodies for detection of the primary antioxidant enzymes.
Fourteen commercial antibodies against human antioxidant enzymes were tested on whole cell lysates by Western analysis for specificity and species crossreactivity. All antibodies, except one, recognized pure protein antigen. All four catalase antibodies were of high quality, and they could also recognize the catalases from rat, mouse, dog, and hamster cells. Two CuZnSOD antibodies were very specific for CuZnSOD protein. They could also crossreact with CuZnSOD from rat, mouse, and hamster cells, but not from dog cells. All five MnSOD antibodies detected only very high levels of MnSOD. We believe that they could not properly be used in immunohistochemistry. Three GPX antibodies could not detect the specific GPX band from cell lysates. We believe that it is difficult to use these GPX antibodies in both Western blotting and immunohistochemistry.
Yan
T
T
Radiation Research Laboratory, Medical Laboratories, The University of Iowa, Iowa City 52242, USA.
Jiang
X
X
Zhang
H J
HJ
Li
S
S
Oberley
L W
LW
D000906:Antibodies / Q000276:immunology*; D000975:Antioxidants / Q000032:analysis*; D002374:Catalase / Q000032:analysis; D004798:Enzymes / Q000032:analysis* / Q000276:immunology; D005979:Glutathione Peroxidase / Q000032:analysis; D006801:Humans; D015151:Immunoblotting; D013482:Superoxide Dismutase / Q000032:analysis; D014407:Tumor Cells, Cultured
D000906:Antibodies; D000975:Antioxidants; D004798:Enzymes; D002374:Catalase; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase