Production and characterization of polyclonal antibodies against Awnt-1, the axolotl homologue of the proto-oncogene product Wnt-1.
The proto-oncogene Wnt-1 (int-1) is activated by mouse mammary tumor virus retroviral insertion and contributes to the formation of mammary gland tumors in mice. During early development, it is expressed in small groups of cells in the developing central nervous system of fish, amphibians and mice. In the Ambystoma mexicanum (axolotl) embryo, the expression profile of Wnt-1 is biphasic. We have raised polyclonal antibodies against a synthetic peptide corresponding to a surface-exposed epitope of Awnt-1. The antibodies react with Awnt-1 synthesized in bacteria as determined by Western blot analysis. They immunoprecipitate the 40 kD Awnt-1 polypeptide translated in vitro from its corresponding cDNA and immunoprecipitation can be efficiently blocked by the immunizing peptide. These data show that the polyclonal anti-peptide antibodies specifically recognize Awnt-1 in its denatured and native conformation.
Busse
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Centre de recherche en cancérologie, Université Laval, L'Hôtel-Dieu de Québec, Canada.
Séguin
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D000558:Ambystoma mexicanum; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000917:Antibody Formation; D015153:Blotting, Western; D000939:Epitopes / Q000276:immunology; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000276:immunology*; D011233:Precipitin Tests; D011487:Protein Conformation; D011505:Protein-Tyrosine Kinases / Q000737:chemistry / Q000276:immunology; D011518:Proto-Oncogene Proteins / Q000737:chemistry / Q000276:immunology*; D014158:Transcription, Genetic; D051153:Wnt Proteins; D051155:Wnt1 Protein; D029961:Zebrafish Proteins
D000906:Antibodies; D000939:Epitopes; D009842:Oligopeptides; D011518:Proto-Oncogene Proteins; D051153:Wnt Proteins; D051155:Wnt1 Protein; D029961:Zebrafish Proteins; D011505:Protein-Tyrosine Kinases