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Generation of a panel of antibodies against proteins encoded on human chromosome 21
BACKGROUND: Down 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 an...
Autores principales: | , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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BioMed Central
2010
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2936279/ https://www.ncbi.nlm.nih.gov/pubmed/20727138 http://dx.doi.org/10.1186/1477-5751-9-7 |
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author | Wiseman, Frances K Sheppard, Olivia Linehan, Jacqueline M Brandner, Sebastian Tybulewicz, Victor LJ Fisher, Elizabeth MC |
author_facet | Wiseman, Frances K Sheppard, Olivia Linehan, Jacqueline M Brandner, Sebastian Tybulewicz, Victor LJ Fisher, Elizabeth MC |
author_sort | Wiseman, Frances K |
collection | PubMed |
description | BACKGROUND: Down 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 RESULTS: We 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. CONCLUSIONS: Although 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. |
format | Text |
id | pubmed-2936279 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-29362792010-09-10 Generation of a panel of antibodies against proteins encoded on human chromosome 21 Wiseman, Frances K Sheppard, Olivia Linehan, Jacqueline M Brandner, Sebastian Tybulewicz, Victor LJ Fisher, Elizabeth MC J Negat Results Biomed Research BACKGROUND: Down 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 RESULTS: We 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. CONCLUSIONS: Although 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. BioMed Central 2010-08-20 /pmc/articles/PMC2936279/ /pubmed/20727138 http://dx.doi.org/10.1186/1477-5751-9-7 Text en Copyright ©2010 Wiseman et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Wiseman, Frances K Sheppard, Olivia Linehan, Jacqueline M Brandner, Sebastian Tybulewicz, Victor LJ Fisher, Elizabeth MC Generation of a panel of antibodies against proteins encoded on human chromosome 21 |
title | Generation of a panel of antibodies against proteins encoded on human chromosome 21 |
title_full | Generation of a panel of antibodies against proteins encoded on human chromosome 21 |
title_fullStr | Generation of a panel of antibodies against proteins encoded on human chromosome 21 |
title_full_unstemmed | Generation of a panel of antibodies against proteins encoded on human chromosome 21 |
title_short | Generation of a panel of antibodies against proteins encoded on human chromosome 21 |
title_sort | generation of a panel of antibodies against proteins encoded on human chromosome 21 |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2936279/ https://www.ncbi.nlm.nih.gov/pubmed/20727138 http://dx.doi.org/10.1186/1477-5751-9-7 |
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