The Isolation and Characterization of Bovine Factor VIII (Antihemophilic Factor)

Heparin sodium salt (Grade I) and e-amino-n-caproic acid were purchased from Sigma.Benzamidine hydrochloride was obtained from Aldrich.Barium sulfate (x-ray grade) was purchased from Merck (U.S.), and kaolin N. F. was obtained from J. T. Baker Chemical Co.Bentonite was a product from Prolabo, Paris, France.Sodium barbital was purchased from Mallinckrodt Chemical Works.Polyethylene glycol (Carbowax 6000) was obtained from Union Carbide Corp., and imidazole, p-alanine, and calcium citrate (Ca3(C6H50& '4 HzO) were obtained through Matheson Coleman and Bell, Cincinnati, Ohio. Cellulose powder was WhatmanCFll fibrous powder, Reeve Angel, Clifton, N. J.The final step of gel filtration was carried out with Agarose A-15m or A-50m supplied by Bio-Rad, Richmond, Calif.Methyl-a-n-glucopyranoside (B grade) was obtained from Calbiochem.Asolecithin, purchased from Associated Concentrates Lecithin, Woodside, N. Y., was used as a platelet substitute.Dri Film silicone (SC 87) was a product from General Electric, Waterford, N. Y., and all glassware was coated with this material unless otherwise noted.Human fibrinogen was purchased from Warner-Chilcott, Morris Plains, N. J. Bovine thrombin employed for the fibrinogen assays was