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JHC exPRESS: First Published January 22, 2008. doi:10.1369/jhc.7A7350.2008
Copyright © Histochemical Society, Inc.

A more recent version of this article appeared on April 1, 2008.
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Articles

Specificity of Four Monoclonal Anti-Nkx6-1 Antibodies

Rasmus Klinck 1, Palle Serup 1, Ole D. Madsen 1 and Mette C. Jørgensen 1*

1 Department of Developmental Biology, Hagedorn Research Institute, Gentofte, Denmark

* To whom correspondence should be addressed. E-mail: mecj{at}hagedorn.dk.

Submitted on September 7, 2007
Accepted on 8 January 2008


  Abstract
The homeodomain transcription factor Nkx6-1 is essential for proper motor neuron development and development of insulin-producing pancreatic {beta}-cells. Nkx6-1 is closely related to Nkx6-2 and Nkx6-3 and all three are expressed in the developing CNS and in the developing foregut. Immunohistochemical detection of protein expression is an important tool for description of the temporal differences in expression patterns. When several gene family members like the Nkx6 factors have overlapping or juxtaposed expression domains, there is an elevated risk of cross-reactivity not being recognized and it is therefore crucial to determine the specificities of antibodies against such targets. In this study, we have determined the epitope consensus sequences of four monoclonal antibodies against Nkx6-1 using SPOT membranes and refined the results by combined peptide recognition and blocking assays. We show that two of the monoclonal anti-Nkx6-1 antibodies specifically recognize Nkx6-1 and do not cross react to Nkx6-2 and Nkx6-3. The other two monoclonal anti-Nkx6-1 antibodies are specific to Nkx6-1 in mice, but do not recognize Nkx6-1 in chicken and human.

Key Words: Nkx6-1, pancreas, neural tube, development, antibody specificity


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