Detection of human, rat and mouse adenosine deaminase by immunochemical and immunomorphologic methods using antiserum to calf enzyme

Abstract

A commercial preparation of calf adenosine deaminase (calf ADA) was further purified by affinity chromatograohy and used for immunization of rabbits. The resulting anti-calf-ADA sera reacted by immunodiffusion with both calf and human ADA, and precipitated about 90% of radiolabeled enzyme isolated from human thymus tissue. Moreover, ADA activity was detected in the pellets formed by immunoprecipitation of unlabeled human enzyme by anti-calf-ADA sera. These antisera were successfully used for the immunomorphologic localization of ADA in human thymus tissue and in lymphoid cell preparations. The anti-calf-ADA sera could also be used for the immunofluorescent detection of enzyme in rat and mouse thymocytes. The utilization of anti-calf-ADA serum for immunochemical and immunomorphologic detection of enzyme provides a valuable and sensitive reagent for the identification of ADA-positive cells in humans and several other species.