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Anal Biochem. 2011 Mar 15;410(2):313-5. doi: 10.1016/j.ab.2010.12.009. Epub 2010 Dec 10.

An assay to measure the affinity of proteins for microtubules by quantitative fluorescent microscopy.

Author information

1
Department of Biochemistry, University of Washington, Seattle, WA 98115, USA.

Abstract

We report a fluorescence-based assay for measuring the affinity of microtubule binding proteins for microtubules. The affinity of any fluorescently tagged protein for taxol-stabilized microtubules can be measured with this assay. We describe the assay and provide a detailed protocol. Using this assay, we found that the affinity of the Dam1 complex for microtubules is decreased by the presence of free unpolymerized tubulin and is sensitive to the salt concentration in the binding buffer. These effects may account for the previous differences in binding affinities reported.

PMID:
21146493
PMCID:
PMC3035983
DOI:
10.1016/j.ab.2010.12.009
[Indexed for MEDLINE]
Free PMC Article

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