Journal
ANALYTICAL BIOCHEMISTRY
Volume 410, Issue 2, Pages 313-315Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2010.12.009
Keywords
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Funding
- NIGMS NIH HHS [R01 GM040506-20, R01 GM040506] Funding Source: Medline
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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. (C) 2010 Elsevier Inc. All rights reserved.
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