title: Characterization of anti-tissue factor antibody and its use in immunoaffinity purification of human tissue factor.
abstract: Human brain tissue factor apoprotein was recently purified to homogeneity in this laboratory by affinity chromatography utilizing factor VII bound to immobilized anti-factor VII (Anal Biochem 165, 365-370, 1987). A potent polyclonal anti-tissue factor antibody has now been raised to the purified apoprotein. Immunostaining of purified tissue factor with this polyclonal IgG yielded a single major band with an apparent molecular weight of 47,000, which corresponds to the mobility on SDS-PAGE of tissue factor apoprotein. Immunostaining of a crude Triton extract of brain tissue yielded two additional bands, with apparent molecular weights of 54,000 and 40,000 Da. An anti-tissue factor IgG was coupled to Affi-Gel-15 to prepare an immunoadsorbant column. The two additional proteins in the crude Triton extract recognized by Western blotting did not bind to the column. This permitted its use to develop a simple, efficient technique for purification of human tissue factor apoprotein.
authors.affiliation: Department of Medicine, University of California, San Diego, La Jolla 92093.
authors.forename: L V
authors.initials: LV
authors.lastname: Rao
mesh_terms: D000906:Antibodies; D001059:Apoproteins / Q000276:immunology / Q000302:isolation & purification; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007074:Immunoglobulin G; D008970:Molecular Weight; D012967:Sodium Dodecyl Sulfate; D013925:Thromboplastin / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D000906:Antibodies; D001059:Apoproteins; D007074:Immunoglobulin G; D012967:Sodium Dodecyl Sulfate; D013925:Thromboplastin