3.7. Tubulin Polymerization Inhibition Assay

The microtubule polymerization assay was carried out in 96-well plates at 37 °C, with 1 mg/mL bovine microtubule-associated protein (MAP)-rich tubulin (Cytoskeleton) and the indicated test compound in PEM buffer [80 mM PIPES (pH 6.8), 1 mM EGTA, and 1 mM MgCl₂] containing 1 mM GTP. To measure the enhancement of polymerization, the microtubule polymerization assay was carried out in 96-well plates at 25 °C, with 1 mg/mL bovine MAP-rich tubulin and the indicated test compound in PEM buffer. Tubulin polymerization was monitored by changes in absorbance at 340 nm [19].