Specificity of antibodies against rodent transforming growth factor-alpha protein.
We found that the immunohistochemical distribution of TGF-alpha varied in rodent tissues depending on the antibody used, suggesting that the specificity of anti-TGF-alpha antibodies differs significantly. To address this issue, we compared the specificity of two representative antibodies that have been widely used to detect rodent TGF-alpha. In a competition study, the antibodies were preincubated with an excess of synthetic rat TGF-alpha 34-50 and were used for staining of rat and mouse kidneys and/or uterus. The results revealed that one of the antibodies, anti-rat TGF-alpha polyclonal antibody, was neutralized by the peptide, whereas the other, anti-human TGF-alpha monoclonal antibody, was not absorbed by the peptide up to an excess of 100-fold. Western blotting analysis showed that the anti-rat TGF-alpha polyclonal antibody recognized both human and rat purified TGF-alpha. However, the anti-human TGF-alpha monoclonal antibody did not detect purified rat TGF-alpha, although the antibody reacted with mouse proteins other than TGF-alpha from kidneys and uterus, purified human TGF-alpha, and mouse carbonic anhydrase II. These data indicate that the anti-human TGF-alpha monoclonal antibody does not recognize rodent TGF-alpha under our experimental conditions and suggest that distribution of TGF-alpha in rodent tissues may need to be reexamined.
Aoyama
H
H
Laboratory of Experimental Pathology, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709, USA.
Kato
H
H
Dixon
D
D
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D003429:Cross Reactions; D005260:Female; D006801:Humans; D051379:Mice; D051381:Rats; D011916:Rats, Inbred F344; D012377:Rodentia; D016211:Transforming Growth Factor alpha / Q000276:immunology*
D000906:Antibodies; D016211:Transforming Growth Factor alpha