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Tubulin polymerization assay

ML Mathias Lundberg
ÅM Åse Mattsson
KR Kathrin Reiser
AH Arne Holmgren
SC Sophie Curbo
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Tubulin polymerization was determined in vitro with a tubulin polymerization assay kit (Cytoskeleton, Inc) according to the manufacturer’s protocol. Briefly, the assay plate was pre-warmed to 37 °C before 5 µl of a 10 × stock solution of the drug of interest or control was added to each well. The plate was then incubated at 37 °C for 2 min before 45 µl of a tubulin solution was added to each well. Polymerization of microtubules was then immediately initiated and measured as enhancement of fluorescence. Readings were done on an Infinite M200 (Tecan) using excitation at 360 nm and emission at 450 nm. Data was collected with one-minute intervals for 60 min and analyzed using Magellan software. All data points were in duplicate and shown as arbitrary fluorescence units (AFU). The data is based on three separate experiments.

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