Immunolabeling of Nuclei/Chromosomes in Arabidopsis thaliana.
Immunolabeling of Nuclei/Chromosomes in Arabidopsis thaliana.
复制标题
拟南芥细胞核/染色体的免疫标记
DOI：
10.1007/978-1-4939-3142-2_10
复制
发表时间：
2016
影响因子：
--
通讯作者：
Lermontova
中科院分区：
文献类型：
--
作者：
Sandmann;Lermontova
细胞周期是细胞生长和有丝分裂或减数分裂的一系列复杂事件。有丝分裂产生两个相同的子细胞，而减数分裂产生配子作为有性生殖的先决条件。为了研究参与细胞周期调控和过程的蛋白质的定位和动力学，可以对融合到荧光标签上的蛋白质进行生命细胞成像。然而，在某些情况下，这种方法不能应用，例如，由于荧光强度低，快速漂白或重组蛋白通过蛋白酶体途径降解。相反，用蛋白质特异性抗体进行免疫标记是分析完整细胞的一种有用的方法。另外，免疫标记也可以应用于特定细胞周期阶段（G1， S和G2）或不同内多倍体水平的分离和/或流动分类的细胞核。本章详细介绍了间接免疫标记方案，以分析拟南芥细胞周期特异性蛋白的亚细胞定位和分布。
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 inArabidopsis thaliana.
影响因子：
7.2
作者：
Lermontova, Inna;Koroleva, Olga;Schubert, Ingo
通讯作者：
Schubert, Ingo