Generating Taxol-Stabilized Microtubules

tubulin protein polymerization mix

Tubulin polymerization mixes containing Alexa 488-labeled tubulin (left), Alexa 594-labeled tubulin (center), and Cy5-labeled tubulin (right) at a 1:4 ratio with unlabeled tubulin.

Overview

Taxol represents a unique class of tubulin inhibitors in that it stabilizes microtubules from depolymerization. It can therefore be utilized in a lab setting to protect microtubules from depolymerization at room temperature. While tubulin protein can be polymerized in the presence of taxol, this has been shown to alter the structure of the subsequent microtubules. We therefore recommend adding taxol to already polymerized microtubules to minimize these effects. Note that when using taxol-stabilized microtubules, taxol must be included in all subsequent buffers. As taxol is highly insoluble in aqueous solutions, it is often necessary to also include DMSO at ~10% to prevent taxol precipitation.

Taxol-stabilized microtubules are relatively long and flexible. To generate short, rigid microtubules, see Generating GMPCPP-Stabilized Microtubules.

Protocol