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在整个细胞群在单细胞水平上研究细胞周期蛋白B的蛋白水解

JoVE Journal
Biologia

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JoVE Journal Biologia

Studying Proteolysis of Cyclin B at the Single Cell Level in Whole Cell Populations

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在整个细胞群在单细胞水平上研究细胞周期蛋白B的蛋白水解

DOI:

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10:54 min

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September 17, 2012

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Dominik Schnerch, Marie Follo, Julia Felthaus, Monika Engelhardt, Ralph Wäsch

¹Department of Hematology, Oncology and Stem Cell Transplantation,University Medical Center Freiburg

Capítulos

00:05Título
01:13Seeding of U2OS-based Cyclin B-SNAP Reporter Cells
03:38Staining of Reporter Cells with SNAP Substrate
05:34Measurement of Fluorescence Intensity
07:07Analysis of Proteolytic Profiles using Scan^R
09:15Results: Cyclin B Kinetics during Mitosis
10:31Conclusion

Summary

Tadução automática

中期向后期过渡被触发，通过后期促进复合物（APC / C）依赖的泛素化和随后的破坏细胞周期蛋白B.在这里，我们建立了一个系统，脉冲追踪标签，可以监控在整个细胞群的细胞周期蛋白B蛋白水解和有利于检测干扰的有丝分裂关卡。

Tags

Proteolysis Cyclin B Single Cell Level Whole Cell Populations Chromosome Distribution Genetic Stability Spindle Assembly Checkpoint (SAC)Mitotic Surveillance Mechanism APC/C Subunit Cdc20 Securin Mitotic Exit Chromosome Separation Improper Chromosomal Attachment SAC Signaling APC/CCdc20 Microtubules Spindle Disruption Artificial Effects

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