The Isolation and Characterization of Bovine Factor VIII
نویسنده
چکیده
Bovine Factor VIII (antihemophilic factor) has been purified approximately lO,OOO-fold with an over-all yield of 10 to 15%. The purification procedure involves BaS04, kaolin, and bentonite adsorption to remove contaminants, ethanol, polyethylene glycol, and /Y-alanine fractionation, calcium citrate-cellulose chromatography, concanavalin A precipitation, and an Agarose gel filtration step. The final product is homogeneous when examined by gel filtration, density gradient centrifugation, and zone electrophoresis. It also shows a single precipitin line when subjected to immunoelectrophoresis employing a specific rabbit antibody against Factor VIII. However, when examined by sedimentation equilibrium, the preparation is physically heterogeneous, apparently due to substantial aggregation of the protein. In these experiments, the smallest species which could be calculated has a molecular weight of approximately 1.1 million. After reduction in Z-mercaptoethanol, the protein shows one band in polyacrylamide gel electrophoresis and zone electrophoresis and a reduction in size. The smallest species of the reduced protein has a molecular weight of about 85,000. The protein contains approximately 11% hexose, 2 % sialic acid, 7 % hexosamine, and no lipid. Other general properties of this protein including its amino acid composition are also reported.
منابع مشابه
Isolation and characterization of a bovine herpesvirus 3 From cases of calf pneumonia in Iran
متن کامل
Development and Cytogenetic Characterization of a Continuous Bovine Kidney Cell Line (IRKHBK) and Evaluation its Susceptibility to some Viruses
In this syudy a continuous bovine kidney cell line derived from a primary bovine kidney cells was established for the first time in Iran. The cells were originating from two-day-old normal male calf of Holstein breed. The cell cultures were continuously passaged following complete proliferation of primary cells. The specific properties or characteristics of the cell were defined using cytogenet...
متن کاملIsolation and Characterization of Human Factor VIII
Factor VIII (antihemophilic factor) has been purified approximately 500.fold from the cryoprecipitate fraction of human plasma. The isolation procedure involves adsorption of contaminants with Al(OH),, column chromatography on tricalcium citrate-cellulose, precipitation with concanavalin A, and an agarose gel filtration step. The linal product is homogeneous when examined by zone electrophoresi...
متن کاملIsolation of bovine spermatogonial cells and co-culture with prepubertal sertoli cells in the presence of colony stimulating factor-1
BACKGROUND: Spermatogonial stem cells (SSCs) are infrequentself-renewing cells among the type A spermatogoniawithin the seminiferous tubules and are the basis of spermatogenesisin mammalian testis. An adequate number of SSCs is aprimary requirement for the study of their behavior, regulation, andfurther biomanipulation. OBJECTIVES: In this paper, we studiedthe development of the primary co-cult...
متن کاملFACTOR V AND VIII INHIBITOR IN PATIENTS WITH COMBINED FACTOR V AND VIII DEFICIENCY
Patients with coagulation factor(s) deficiency who use coagulation therapy are susceptible to forming inhibitors against coagulation factor(s). In this survey we detected factor V and VIII inhibitor in ten patients with combined deficiency of factors V and VIII from north east of Iran (Khorassan province). It was revealed in our survey that eight patients had both factor V and factor VIII i...
متن کامل