Nuclear proteome: Isolation of intact nuclei, extraction of nuclear proteins, and 2-DE analysis
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Title |
Nuclear proteome: Isolation of intact nuclei, extraction of nuclear proteins, and 2-DE analysis
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Creator |
Pandey, Aarti
Chakraborty, Subhra Chakraborty, Niranjan |
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Subject |
Cellular contaminants
Nuclear proteins DAPI staining Marker enzymes Organelle proteome |
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Description |
Accepted date: 31 October 2017
Proteome profiling aims to unravel the mystery of biological complexity encoded by the genome. The successful proteome profiling largely depends upon analytical approaches because single-step proteome characterization of eukaryotic cells is difficult due to the large number of proteins expressed and their complex physiochemical properties. Organellar proteomics helps in identifying a refined set of proteins by pinpointing certain activities to specific organelles, thereby increasing our knowledge of cellular processes. The reliability of a plant organelle proteome is intimately dependent on the purity of the organelle preparation. Methodological improvements in sample handling, organelle fractionation, and protein extraction are therefore crucial to plant subcellular proteomics. The nuclear proteins are organized into complex regulatory networks and perform varied cellular functions. Therefore, characterization of the nuclear proteome is an important step toward accumulating knowledge about regulation of gene expression and function. In this chapter, we present methods for the isolation of nuclei, purification of nuclear proteins, and proteome profiling that have been adapted for proteomic characterization of economically important crop species, such as chickpea. This research work was supported by grants from the Department of Biotechnology (DBT), Ministry of Science and Technology, India and the National Institute of Plant Genome Research to N. C. Aarti Pandey is recipient of the Post-doctoral Fellowship from the National Institute of Plant Genome Research. |
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Date |
2017-11-02T08:52:56Z
2017-11-02T08:52:56Z 2018 |
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Type |
Book chapter
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Identifier |
Methods in Molecular Biology, 1696: 41-55
978-1-4939-7411-5 http://223.31.159.10:8080/jspui/handle/123456789/793 http://www.springerprotocols.com/Abstract/doi/10.1007/978-1-4939-7411-5_3 10.1007/978-1-4939-7411-5_3 |
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Language |
en_US
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Format |
application/pdf
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Publisher |
Springer Nature
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