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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
 

Purification of Tubulin with Controlled Posttranslational Modifications and Isotypes from Limited Sources by Polymerization-Depolymerization Cycles

Article DOI: 10.3791/61826-v • 07:54 min • November 5th, 2020
November 5th, 2020 •
*1,2, *1,2, 1,2, 1,2
1PSL Research University, CNRS UMR3348, Institut Curie, 2Université Paris-Saclay, CNRS UMR3348, Université Paris Sud
* These authors contributed equally

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This protocol describes tubulin purification from small/medium-scale sources such as cultured cells or single mouse brains, using polymerization and depolymerization cycles. The purified tubulin is enriched in specific isotypes or has specific posttranslational modifications and can be used in in vitro reconstitution assays to study microtubule dynamics and interactions.

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Purification Tubulin Posttranslational Modifications Isotypes Polymerization-Depolymerization Cycles Assembly-competent Tubulin In-vitro Experiments Cell Biology Lab Hands-on Time Suspension Cultures Revive Amplify Confluent Dishes Spinner Bottles Medium Equilibration Inoculate Harvest Cells Centrifugation Resuspend Cells PBS
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