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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

JoVE Journal
Biochimica

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JoVE Journal Biochimica

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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Satish Bodakuntla*^1,2, A.S. Jijumon*^1,2, Carsten Janke², Maria M. Magiera²

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

Capitoli

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

Traduzione automatica

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

Keywords: Tubulin Purification Posttranslational Modifications Isotypes Polymerization-depolymerization Cycles Suspension Cultures Adherent Cell Cultures Cell Harvesting Lysis Buffer PBS-EDTA

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