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Methods Cell Biol. 2013;115:215-29. doi: 10.1016/B978-0-12-407757-7.00014-1.

Measurement of in vitro microtubule polymerization by turbidity and fluorescence.

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1
Program in Physical Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, Maryland, USA.

Abstract

Tubulin polymerization may be conveniently monitored by the increase in turbidity (optical density, or OD) or by the increase in fluorescence intensity of diamidino-phenylindole. The resulting data can be a quantitative measure of microtubule (MT) assembly, but some care is needed in interpretation, especially of OD data. Buffer formulations used for the assembly reaction significantly influence the polymerization, both by altering the critical concentration for polymerization and by altering the exact polymer produced-for example, by increasing the production of sheet polymers in addition to MT. Both the turbidity and the fluorescence methods are useful for demonstrating the effect of MT-stabilizing or -destabilizing additives.

KEYWORDS:

Critical concentration; Fluorescence; Microtubules; Optical density; Polymerization; Tubulin; Turbidity; Turbidity coefficient

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