Unit

UNIT 3.29 Isolation of Microtubules and Microtubule Proteins

  1. J. Avila1,
  2. H. Soares2,
  3. M.L. Fanarraga3,
  4. J.C. Zabala3

Published Online: 1 JUN 2008

DOI: 10.1002/0471143030.cb0329s39

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Avila, J., Soares, H., Fanarraga, M. and Zabala, J. 2008. Isolation of Microtubules and Microtubule Proteins. Current Protocols in Cell Biology. 39:3.29:3.29.1–3.29.28.

Author Information

  1. 1

    Centro de Biología Molecular (CSIC). Facultad de Ciencias, Universidad Autónoma de Madrid, Cantoblanco, Madrid, Spain

  2. 2

    Instituto Gulbenkian de Ciência, 2781-901, Oeiras, and Escola Superior de Tecnologia da Saúde de Lisboa, Lisboa, Portugal

  3. 3

    Departamento de Biología Molecular, Facultad de Medicina, Universidad de Cantabria- IFIMAV, Santander, Spain

Publication History

  1. Published Online: 1 JUN 2008
  2. Published Print: JUN 2008

Abstract

This unit describes various protocols for the isolation and purification of the main constituents of microtubules, chiefly α- and β-tubulin, and the most significant microtubule associated proteins (MAPs), specifically MAP1A, MAP1B, MAP2, and tau. We include a classical isolation method for soluble tubulin heterodimer as the first basic purification protocol. In addition, we show how to analyze the tubulin and MAPs obtained after a phosphocellulose chromatography purification procedure. This unit also details a powerful and simple method to determine the native state of the purified tubulin based on one-dimensional electrophoresis under nondenaturing conditions (unit 6.5). The last protocol describes the application of a new technique that allows visualizing the quality of polymerized microtubules based on atomic force microscopy (AFM). Curr. Protoc. Cell Biol. 39:3.29.1-3.29.28. © 2008 by John Wiley & Sons, Inc.

Keywords:

  • tubulin;
  • microtubule;
  • MAP1A;
  • MAP1B;
  • MAP2;
  • tau;
  • native gel electrophoresis;
  • atomic force microscopy