Abstract
The current protocol describes in the first part the isolation and culture of neural stem and progenitor cells. In the second part, the two-dimensional gel electrophoresis of neural stem and progenitor cells is explained, consisting of sample preparation, first-dimensional isoelectric focusing, second-dimensional acrylamide gel electrophoresis, protein staining, and image digitalization. The third part concentrates on the software analysis of two-dimensional gel images and the bioinformatical processing.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Brüstle, O., and McKay, R. D. (1996) Neuronal progenitors as tools for cell replacement in the nervous system, Curr. Opin. Neurobiol. 6, 688–695.
Gage, F. H. (2000) Mammalian neural stem cells, Science 287, 1433–1438.
Okano, H. (2002) Stem cell biology of the central nervous system, J. Neurosci. Res. 69, 698–707.
Maurer, M. H., Feldmann, R. E., Jr., Fütterer, C. D., and Kuschinsky, W. (2003) The proteome of neural stem cells from adult rat hippocampus, Proteome Sci. 1, 4.
Link, A., (Ed.) (1999) 2-D proteome analysis protocols, Humana Press, Totowa, N.J.
Maurer, M. H., and Kuschinsky, W. (2007) Proteomics, in Handbook of Neurochemistry and Molecular Neurobiology, vol. 5: Brain Energetics. Integration of Cellular and Molecular Processes. (Lajtha, A., Gibson, G. E., and Dienel, G. A., Eds.) 3 rd ed., pp 737–769, Springer, New York.
Görg, A., Obermaier, C., Boguth, G., Harder, A., Scheibe, B., Wildgruber, R., and Weiss, W. (2000) The current state of two-dimensional electrophoresis with immobilized pH gradients, Electrophoresis 21, 1037–1053.
Rabilloud, T., Vaezzadeh, A. R., Potier, N., Lelong, C., Leize-Wagner, E., and Chevallet, M. (2009) Power and limitations of electrophoretic separations in proteomics strategies, Mass Spectrom. Rev. 28, 816–843.
Berkelman, T., and Stenstedt, T. (2002) 2-D Electrophoresis using immobilized pH gradients. Principles and Methods, 2nd ed., Amersham, Uppsala.
Maurer, M. H. (2006) Software analysis of two-dimensional electrophoretic gels in proteomic experiments, Curr. Bioinformatics. 1, 255–262.
Sheehan, D., and Tyther, R., (Eds.) (2009) Two-Dimensional Electrophoresis Protocols, Vol. 519, Humana Press, New York.
Maurer, M. H. (2004) The path to enlightenment: making sense of genomic and proteomic information, Genomics Proteomics Bioinformatics 2, 123–131.
Acknowledgments
The author was supported by the European Union (EU) within the Seventh Framework Programme (FP7), the German Ministry of Education and Research (BMBF) within the National Genome Research Network (NGFN-2), the German Research Foundation (DFG), the Intramural Research Grant of the Medical Faculty, University of Heidelberg, the Steuben-Schurz Society, and the Estate of Friedrich Fischer.
The author thanks Prof. em. Dr. Wolfgang Kuschinsky for his sustained support for many years, Mrs. Maria Harlacher, Mrs. Tilly Lorenz, and Mrs. Inge Keller for excellent technical support.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Maurer, M.H. (2012). Two-Dimensional Protein Analysis of Neural Stem Cells. In: Karamanos, Y. (eds) Expression Profiling in Neuroscience. Neuromethods, vol 64. Humana Press. https://doi.org/10.1007/978-1-61779-448-3_7
Download citation
DOI: https://doi.org/10.1007/978-1-61779-448-3_7
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-447-6
Online ISBN: 978-1-61779-448-3
eBook Packages: Springer Protocols