Production of antibodies to human factor VIII in the ascitic fluid of C57BL/6J mice.

A method is described for the production of a murine antiserum to human factor VIII, and the quantitation of factor VIII related antigen using this antiserum. C57BL/6J mice were injected intraperitoneally with a mixture of factor VIII and complete Freund's adjuvant. Ascitic fluid developed after repeated innoculations for seven to eight weeks, and was subsequently tapped. Using this antiserum, factor VIII related antigen was quantitated by immunoelectrophoresis and enzyme immunoassay. This method provides a simple and economical way of producing an antiserum to factor VIII, and may also be applicable to other proteins available in submiligram quantities.

beta-Glucuronidase activation of latent aggression-promoting cues in mouse bladder urine.

The present study was designed to investigate the chemical properties of the aggression-promoting cues present in bladder urine of male mice. The results of the first experiment confirmed earlier work by demonstrating the presence of an aggression-promoting chemosignal in bladder urine. In Experiment 2, behavioral assays were separately performed on the organic and aqueous layers of bladder urine obtained by repeated dichloromethane extractions. Only the combined organic layers of the initial three extractions demonstrated behavioral activity. A fourth extraction showed no behavioral activity for both organic and aqueous layers. However, the findings of Experiment 3 showed that incubation of the aqueous layer from the third CH2Cl2 extraction in beta-glucuronidase can free additional aggression-promoting cues into a subsequent CH2Cl2 extraction. It is concluded that two forms of the aggression-promoting chemosignal are present in bladder urine. One is lipophilic and behaviorally active, whereas the other is conjugated, possessing latent chemosignal properties.