Summary
Two-dimensional electrophoresis (2DE) is one of the most effective methods for the reliable separation of proteins in a single gel. In our proteome analyses of human chromosomes, we used two types of 2DE: two-dimensional isoelectric focusing SDS–polyacrylamide gel electrophoresis (2D IEF/SDS–PAGE) and radical-free and highly reduced two-dimensional polyacrylamide gel electrophoresis (RFHR 2D PAGE) together with one-dimensional SDS–polyacrylamide gel electrophoresis (1DE). Experimental details of these gel electrophoresis procedures that have been shown to be effective for human proteome analyses are described in detail.
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Acknowledgments
The authors would like to thank Drs. Sachihiro Matsunaga, Shin’ichiro Kajiyama, and Hideki Takata for their collaboration during the research on chromosome proteomics. We also thank Drs. Hideji Yoshida and Akira Wada for the technical advice of RFHR 2D PAGE. This work was supported by the Special Coordination Funds from the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan, to K. F. and S.U.
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Fukui, K., Uchiyama, S. (2009). 2DE for Proteome Analysis of Human Metaphase Chromosomes. In: Tyther, R., Sheehan, D. (eds) Two-Dimensional Electrophoresis Protocols. Methods in Molecular Biology, vol 519. Humana Press. https://doi.org/10.1007/978-1-59745-281-6_16
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DOI: https://doi.org/10.1007/978-1-59745-281-6_16
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