A commercial fluorescence-based tubulin polymerization assay (Cytoskeleton, Denver, CO; Cat. # BK011P) was used as per manufacturer’s instructions for detection of tubulin polymerization inhibitors. Briefly, porcine tubulin (3 mg/ml tubulin in 80 mM PIPES pH 7.0, 0.5 mM EGTA, 2 mM MgCl2, 1 mM GTP and 10% glycerol) was incubated at 37 °C with either vehicle control (DMSO), with Paclitaxel or antimitotic compound. Each condition was tested in duplicate. Polymerization was measured over time by excitation at 360 nm and emission at 420 nm in a temperature controlled 96-well plate fluorimeter (Envision 2105, Perkin-Elmer, Waltham, MS) equipped with filters for excitation at 340–360 nm and emission at 420–450 nm.
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