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Generation and Characterization of Monoclonal Antibodies against the Transcription Factor Nkx6.1

Authors :
Ole D. Madsen
Mette C. Jørgensen
Rasmus Klinck
Jacob Hecksher-Sørensen
Stefan Zahn
Palle Serup
Inger Lund Pedersen
Source :
Journal of Histochemistry & Cytochemistry. 54:567-574
Publication Year :
2006
Publisher :
SAGE Publications, 2006.

Abstract

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 β-cells in the pancreas. Expression of Nkx6.1 can be detected in developing pancreatic epithelium and in adult insulin-producing β-cells, making this transcription factor a unique β-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/κ subtype. When applied for immunohistochemistry on frozen sections of adult mouse pancreas, monoclonal antibodies stain specifically the β-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. (J Histochem Cytochem 54:567-574, 2006)

Details

ISSN :
15515044 and 00221554
Volume :
54
Database :
OpenAIRE
Journal :
Journal of Histochemistry & Cytochemistry
Accession number :
edsair.doi.dedup.....785315ab1711b0df96af95e41b1e2160
Full Text :
https://doi.org/10.1369/jhc.5a6827.2006