Abstract
Naked2 (NKD2) is a member of the Naked family and negatively regulates canonical Wnt signaling. NKD2 may play a role in embryo development and tumor formation by affecting Wnt signaling. In the present study, we describe the establishment of a monoclonal antibody against NKD2 (anti-NKD2 MAb) through the hybridoma method. The purified anti-NKD2 MAb measured a titer of 2.56 × 10(5) against NKD2 by indirect ELISA. Western blot analysis, immunoprecipitation, and confocal microscope showed that the anti-NKD2 MAb can specifically combine NKD2 protein in SW480 and LOVO cells. Competitive inhibition assays of Western blot and indirect ELISA showed that the anti-NKD2 MAb can be blocked with NKD2(1-217) protein. The anti-NKD2 MAb would be helpful for further studies on the structure activity relationship, protein detecting, and cell-signaling pathway of NKD2.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Proteins, Signal Transducing
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Animals
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Antibodies, Monoclonal / biosynthesis
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Antibodies, Monoclonal / immunology*
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Antibodies, Monoclonal / isolation & purification
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Antibody Formation
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Antibody Specificity
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Blotting, Western
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Calcium-Binding Proteins
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Carrier Proteins / immunology*
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Carrier Proteins / isolation & purification
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Carrier Proteins / metabolism
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Colorectal Neoplasms / immunology*
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Colorectal Neoplasms / metabolism
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Enzyme-Linked Immunosorbent Assay
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Female
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Flow Cytometry
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Humans
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Hybridomas / immunology*
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Immunization
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Immunoprecipitation
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Mice
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Mice, Inbred BALB C
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Microscopy, Confocal
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Recombinant Proteins
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Tumor Cells, Cultured
Substances
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Adaptor Proteins, Signal Transducing
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Antibodies, Monoclonal
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Calcium-Binding Proteins
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Carrier Proteins
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NKD2 protein, human
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Recombinant Proteins