Plasminogen activator activity was determined in stimulated normal rabbit tears, and was identified as being due to urokinase using specific antibodies. The activator activity is about 100-fold higher in rabbit tears than in human tears. The molecular weight of the activator was determined by fibrin zymography to be 40-50 kDa. However, a multiband pattern of urokinase-type antigen ranging in molecular weight from 60-70 kDa to 200 kDa was detected by immunoblotting. The 60-70 kDa protein was resistant to reduction. The antigen concentration is more than 100-fold higher than the concentration expected from the activity measurement, indicating that most of the urokinase in rabbit tears is in inactive form, such as in inhibitor or receptor complexes.