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通过聚合-去聚合周期，通过有限来源的受控后翻译修改和异种型对图布林进行净化
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Biochemistry
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JoVE Journal Biochemistry
Purification of Tubulin with Controlled Posttranslational Modifications and Isotypes from Limited Sources by Polymerization-Depolymerization Cycles
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通过聚合-去聚合周期，通过有限来源的受控后翻译修改和异种型对图布林进行净化

DOI:
10.3791/61826-v

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07:54 min

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November 05, 2020

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, , ,
1PSL Research University, CNRS UMR3348,Institut Curie, 2Université Paris-Saclay, CNRS UMR3348,Université Paris Sud

Chapters

  • 00:05Introduction
  • 00:34Amplification and Lysis of Cells in Suspension Cultures
  • 02:08Amplification and Lysis of Adherent Cells and Mouse Brain Tissue Lysis
  • 03:46Sample Preparation for Analysis
  • 04:10Clarification, Polymerization, and Depolymerization of the Lysate
  • 06:27Evaluation, Quantification, and Characterization of Purified Tubulin
  • 07:19Conclusion

Summary

Automatic Translation

Automatic Translation

本协议使用聚合和去聚合周期描述从中小型来源（如培养细胞或单个小鼠大脑）提纯的图布林。纯化的图布林富含特定的同位素或具有特定的翻译后修饰，可用于体外重组测定，以研究微管动力学和相互作用。

Tags

Tubulin PurificationPosttranslational ModificationsIsotypesPolymerization-depolymerization CyclesSuspension CulturesAdherent Cell CulturesCell HarvestingLysis BufferPBS-EDTA

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