Production and characterization of an anti-chicken transferrin.

Several lines of recent evidence have suggested that transferrin plays a significant role in tissue interaction or morphogenesis at early stages of embryo development. In the present study, an anti-chicken transferrin antibody was produced and its basic characteristics were clarified as a basis for use in further studies. An antiserum termed Toraji 3 was raised against chicken transferrin and purified into IgG and ligand-affinity-purified fractions. These three preparations of the antibody gave an intense immunohistochemical signal in visceral yolk sac and developing liver, both of which are known to be major producers of transferrin in early development. In immunoblot analysis, these three preparations detected 70-kDa transferrin, whereas the ligand-affinity-purified preparation showed higher specificity. It was also demonstrated by enzyme-linked immunosorbent assay and immunoblotting that Toraji 3 antibody bound preferentially to chicken transferrin and showed a negligible binding to human transferrin.

[1]  R. Markwald,et al.  Identification of transferrin as one of multiple EDTA‐extractable extracellular proteins involved in early chick heart morphogenesis , 1994, Journal of cellular biochemistry.

[2]  A. Copp,et al.  Exogenous transferrin is taken up and localized by the neurulation-stage mouse embryo in vitro. , 1992, Developmental biology.

[3]  G. Nicolson,et al.  Lung‐derived growth factor that stimulates the growth of lung‐metastasizing tumor cells: Identification as transferrin , 1991, Journal of cellular biochemistry.

[4]  R. MacGillivray,et al.  The nucleotide sequence of rabbit liver transferrin cDNA. , 1991, Biochimica et biophysica acta.

[5]  I. Romslo,et al.  The role of transferrin in the mechanism of cellular iron uptake. , 1990, The Biochemical journal.

[6]  S J Rodda,et al.  Mechanisms of antibody binding to a protein. , 1987, Science.

[7]  R. Lerner,et al.  Chemistry of antibody binding to a protein. , 1987, Science.

[8]  S. Naylor,et al.  Human transferrin: cDNA characterization and chromosomal localization. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[9]  R. Timpl,et al.  Transferrin as a fetal growth factor: acquisition of responsiveness related to embryonic induction. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[10]  J. Sedat,et al.  Fluorescence microscopy: reduced photobleaching of rhodamine and fluorescein protein conjugates by n-propyl gallate. , 1982, Science.

[11]  P. Chambon,et al.  The complete nucleotide sequence of the chicken ovotransferrin mRNA. , 1982, European journal of biochemistry.

[12]  David Barnes,et al.  Serum-free cell culture: a unifying approach , 1980, Cell.

[13]  D. Wallach,et al.  Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. , 1971, Biochemistry.

[14]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[15]  V. Hamburger,et al.  A series of normal stages in the development of the chick embryo , 1951, Journal of morphology.

[16]  P. Aisen,et al.  Iron transport and storage proteins. , 1980, Annual review of biochemistry.