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JHC exPRESS: First Published January 9, 2006. doi:10.1369/jhc.5A6827.2006
Journal of Histochemistry and Cytochemistry
Copyright © 2006 Pedersen et al.
A more recent version of this article appeared on May 1, 2006.
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Articles |
Generation and Characterization of Monoclonal Antibodies against the Transcription Factor Nkx6.1
1 Antibody Core, Beta Cell Biology Consortium, www.betacell.org. (ILP,ODM); Novo Nordisk A/S, Department of Antibody and Cell Technology, Bagsværd, Denmark (ILP,SZ); and Hagedorn Research Institute, Gentofte, Denmark (RK,JH-S,ODM,PS,MCJ)
* To whom correspondence should be addressed. E-mail: pas{at}hagedorn.dk.
Submitted on September 1, 2005
Accepted on 16 December 2005
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Abstract |
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-cells in the pancreas. Expression of Nkx6.1 can be detected in developing pancreatic epithelium and in adult insulin-producing -cells, making this transcription factor a unique -cell marker. For the 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/
subtype. When applied for immunohistochemistry on frozen sections of adult mouse pancreas, the monoclonal antibodies stain specifically the -cells in the endocrine Islets of Langerhans with patterns comparable to that of a previously produced polyclonal rabbit serum. The 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.
Key Words: Nkx6.1, pancreas, neural tube, development, diabetes, antibody
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