It has been suggested that mouse and rat lack adenosine deaminase-complexing protein because in these species exclusively the small molecular weight form of adenosine deaminase (ADA-S) is found. This suggestion is based on the assumption that the adenosine deaminase binding capacity is an inherent functional characteristic of adenosine deaminase-complexing protein. We report on the presence of adenosine deaminase-complexing protein immunoreactivity in mouse and rat determined with a species cross-reactive polyclonal anti-adenosine deaminase-complexing protein serum. In the mouse the tissue and subcellular distribution and the electrophoretic mobility in starch and polyacrylamide gels of the protein correspond with those of adenosine deaminase-complexing protein, but it does not bind the small molecular weight form of adenosine deaminase. Furthermore, in human, mouse, and rat kidney cortex adenosine deaminase and adenosine deaminase-complexing protein did not colocalize by immunohistochemistry. It is suggested that the function of adenosine deaminase-complexing protein is not adenosine deaminase-related.