Chapter title |
Immunolabeling of Nuclei/Chromosomes in Arabidopsis thaliana.
|
---|---|
Chapter number | 10 |
Book title |
Plant Cell Division
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3142-2_10 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3141-5, 978-1-4939-3142-2
|
Authors |
Sandmann, Michael, Fuchs, Joerg, Lermontova, Inna, Michael Sandmann, Joerg Fuchs, Inna Lermontova |
Abstract |
The cell cycle is a complex sequence of events by which cells grow and divide mitotically or meiotically. Mitosis results in the generation of two identical daughter cells, while meiosis generates gametes as a prerequisite for sexual reproduction. To study the localization and dynamics of proteins involved in the regulation and proceeding of the cell cycle, life cell imaging of proteins fused to fluorescent tags can be performed. However, in some cases this approach cannot be applied, e.g., due to low fluorescence intensity, fast bleaching or degradation of recombinant proteins by the proteasome pathway. Instead, immunolabeling with protein-specific antibodies represents a useful approach for the analysis of intact cells. Alternatively, immunolabeling can also be applied to isolated and/or flow-sorted nuclei of particular cell cycle stages (G1, S, and G2) or of different endopolyploidy levels. This chapter details indirect immunolabeling protocols to analyze the subcellular localization and distribution of cell cycle-specific proteins in Arabidopsis thaliana. |
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