I'm running a teaching lab where students are adding taxol dissolved in DMSO to a reaction of tubulin, ddH2O, DMSO, and a buffer (PIPES, 1M MgSO4, water, 0.5 M EGTA, pH adjusted to 6.9, and GTP sodium salt hydrated with dd-water). Whenever I add the taxol , a white precipitate forms. In the absence of taxol there's no precipitate. Any idea why?
Dear Aisling Brady ,
I think probably your students add the taxol solution to the tubulin all at once because this can lead to precipitation of tubulin. To solve this problem, your students can try adding the taxol solution step by step. For example, if they intend to polymerize 2 mg/ml tubulin, they can use 2 µM, 20 µM, and 200 µM steps.
I attached the related protocol for you.
I hope this could be helpful.
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