I would like to perform the microtubule polymerization assay in MCF10A cells.
I found this protocol in one paper, which is pretty straight forward:
Cells plated in glass coverslips were transferred to ice cold medium and incubated 1 h on ice to depolymerize microtubules. Cells were then incubated with medium at 37 uC for 30 s to allow microtubules to polymerize and fixed immediately in ice-cold methanol for 10 min. Cells were stained for microtubules and centrioles (as described above) and microtubule number was quantified manually. Images of cells used in the analysis were acquired from 2 independent experiments.
However, I think some people do this assay with nocodazole pre-treatment. Does anyone have any experience with this assay that can give me a more detailed protocol?