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 submilligram quantities.