I have a problem because when I add doxycycline (in distilled water, DMSO, methanol or PBS according to different protocol) at the doses (100-300) µm/ml to culture media (DMEM), it precipitates (sedimentation forms). I read different articles and I really appreciate your help because I did not find a precise answer. I am very grateful if you response to my question.
Hi,
In our lab we prepare stock solutions of DOX with a concentration of 2 mg/ml (in H2O) - we use it 1:1000 in cell culture (e.g. add 50µl to 50 ml of DMEM) for the induction of Tet-inducible promotors...
Your concentration might be too high?!
Hi Samaneh,
I prepare 10mg/ml stock of Dox in Water and use it a final concentration of 1-5ug/ml cell culture medium like DMEM. Your solvent of choice will depend on the kind of salt you purchase. Usually, sodium salts are soluble in water and others are soluble in EtOH/DMSO etc.
With DMSO, you can frequently observe sediments/precipitation if the concentration of stock solution is too high. But if you use water, it should be fine.
Hope this helps. Good luck!
Utkarsh
Samaneh,
Please clarify what you mean by 100-300 µm/ml concentration. Is it your stock solution or the final concentration in the medium?
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