Single step purification of platelet factor XIII using an immobilized factor XIII a-subunit monoclonal antibody.
To facilitate the analysis of the catalytic subunit of factor XIII we have developed a method for the immunoaffinity purification of this protein from platelets. This method employs a monoclonal antibody that binds to the a-subunit of factor XIII. The anti-factor XIII antibody was immobilized on agarose and then incubated with platelet lysate. Subsequently factor XIII was isolated from the platelet lysate in a single step with a 41% yield as measured by enzyme assay. The purified platelet factor XIII appeared nearly homogeneous when analyzed by polyacrylamide electrophoresis and by immunoblotting with another factor XIII monoclonal antibody.
Massachusetts General Hospital, Boston.
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001792:Blood Platelets / Q000737:chemistry*; D015153:Blotting, Western; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005176:Factor XIII / Q000276:immunology / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D007163:Immunosorbent Techniques; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D005176:Factor XIII