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αβ-Tubulin and Microtubule-Binding Assays

  • Jaime N. Campbell
  • Kevin C. SlepEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 777)

Abstract

Dynamic instability is a hallmark of the microtubule cytoskeleton. In order to regulate microtubule dynamics in vivo, a varied host of microtubule-associated proteins are mobilized to promote microtubule nucleation, growth, stabilization, catastrophe, depolymerization, rescue, and severing. To confer these various functions, cytoskeletal regulators have highly tuned affinities for tubulin, recognizing the unpolymerized αβ-tubulin heterodimer, dynamic microtubule lattice, stabilized microtubule lattice, or a combination therein. The protocols presented here probe αβ-tubulin and microtubule binding using gel filtration and cosedimentation, respectively.

Key words

Tubulin Tubulin-binding protein Microtubule-associated protein Gel filtration Microtubule cosedimentation assay Dynamic microtubules Taxol 

Notes

Acknowledgments

The authors would like to thank Dr. Stephen Rogers for advice. This work was supported by a Basil O’Connor Research Starter Grant from the March of Dimes.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of Biochemistry and BiophysicsUniversity of North CarolinaChapel HillUSA
  2. 2.Molecular and Cellular Biophysics ProgramUniversity of North CarolinaChapel HillUSA
  3. 3.Department of BiologyUniversity of North CarolinaChapel HillUSA

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