Generation of a panel of antibodies against proteins encoded on human chromosome 21Report as inadecuate




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Journal of Negative Results in BioMedicine

, 9:7

First Online: 20 August 2010Received: 15 March 2010Accepted: 20 August 2010DOI: 10.1186-1477-5751-9-7

Cite this article as: Wiseman, F.K., Sheppard, O., Linehan, J.M. et al. J Negat Results BioMed 2010 9: 7. doi:10.1186-1477-5751-9-7

Abstract

BackgroundDown syndrome DS is caused by trisomy of all or part of chromosome 21. To further understanding of DS we are working with a mouse model, the Tc1 mouse, which carries most of human chromosome 21 in addition to the normal mouse chromosome complement. This mouse is a model for human DS and recapitulates many of the features of the human syndrome such as specific heart defects, and cerebellar neuronal loss. The Tc1 mouse is mosaic for the human chromosome such that not all cells in the model carry it. Thus to help our investigations we aimed to develop a method to identify cells that carry human chromosome 21 in the Tc1 mouse. To this end, we have generated a panel of antibodies raised against proteins encoded by genes on human chromosome 21 that are known to be expressed in the adult brain of Tc1 mice

ResultsWe attempted to generate human specific antibodies against proteins encoded by human chromosome 21. We selected proteins that are expressed in the adult brain of Tc1 mice and contain regions of moderate-low homology with the mouse ortholog. We produced antibodies to seven human chromosome 21 encoded proteins. Of these, we successfully generated three antibodies that preferentially recognise human compared with mouse SOD1 and RRP1 proteins on western blots. However, these antibodies did not specifically label cells which carry a freely segregating copy of Hsa21 in the brains of our Tc1 mouse model of DS.

ConclusionsAlthough we have successfully isolated new antibodies to SOD1 and RRP1 for use on western blots, in our hands these antibodies have not been successfully used for immunohistochemistry studies. These antibodies are freely available to other researchers. Our data high-light the technical difficulty of producing species-specific antibodies for both western blotting and immunohistochemistry.

Electronic supplementary materialThe online version of this article doi:10.1186-1477-5751-9-7 contains supplementary material, which is available to authorized users.

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Author: Frances K Wiseman - Olivia Sheppard - Jacqueline M Linehan - Sebastian Brandner - Victor LJ Tybulewicz - Elizabeth MC Fi

Source: https://link.springer.com/







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