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Publication : Generation and characterization of monoclonal antibodies against the transcription factor Nkx6.1.

First Author  Pedersen IL Year  2006
Journal  J Histochem Cytochem Volume  54
Issue  5 Pages  567-74
PubMed ID  16401696 Mgi Jnum  J:226237
Mgi Id  MGI:5696602 Doi  10.1369/jhc.5A6827.2006
Citation  Pedersen IL, et al. (2006) Generation and characterization of monoclonal antibodies against the transcription factor Nkx6.1. J Histochem Cytochem 54(5):567-74
abstractText  We present the generation of a panel of monoclonal antibodies (F55A10, F55A12, F64A6B4, and F65A2) against the homeodomain transcription factor Nkx6.1, one of the essential transcription factors that regulates the multistep differentiation process of precursor cells into endocrine beta-cells in the pancreas. Expression of Nkx6.1 can be detected in developing pancreatic epithelium and in adult insulin-producing beta-cells, making this transcription factor a unique beta-cell marker. For production of monoclonal antibodies, RBF mice were immunized with a GST-Nkx6.1 fusion protein containing a 66-amino acid C-terminal fragment of rat Nkx6.1. Four clones were established as stable hybridoma cell lines and the produced antibodies were of the mouse IgG1/kappa subtype. When applied for immunohistochemistry on frozen sections of adult mouse pancreas, monoclonal antibodies stain specifically the beta-cells in the endocrine islets of Langerhans with patterns comparable to that of a previously produced polyclonal rabbit serum. Monoclonal antibodies can be divided into two groups that appear to recognize different epitopes, as determined by competition ELISA. The presented antibodies are useful tools for the further characterization of the role and function of Nkx6.1 in pancreatic development, especially for use in double-labeling experiments with existing polyclonal rabbit antibodies.
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