Dear all,
I am giving this drug which is 16.5mM in DMSO and I want to make 0.1mM and 0.33mM of it to add it to the cancer cell line. I know DMSO might have an effect on the cells so I'm planning on adding 10Microliter of it to the 10cm dish. but I am really lost on how to make these different concentrations from the original as I want the concentration of DMSO to be the same in all of my plates.
Thank you in advance for anyone who answers
What is the volume of media you are adding to your dish?
Assuming 10mL of media, in your 10cm dish,
To prepare your 0.1mM concentration, take 60.6uL of 16.5mM drug in 10mL of media (so 9,939.4uL of media and 60.6uL of drug). As a control, I would do the same dilution into another dish, but only with DMSO not containing any drug, as your DMSO control.
Similarly for you 0.33mM, 200uL of drug in 10mL of media.
Your DMSO control should reflect what you have done. Dependant on your cells and assays, it may be sufficent to only use the higher DMSO concentration as your control.
However, 10uL of DMSO in 10mL represents a drug concentration of 0.0165mM.
Hello Delilah,
to add to what Sandra already said: "10uL of DMSO in 10mL represents a drug concentration of 0.0165mM" - treating your cells with 0.1% DMSO (10µl in 10ml) will get you a 6-fold lower concentration than even the lower of your desired concentrations of 0.1 and 0.33mM.
So using 0.6% (for 0.1mM) of DMSO will quite likely have some toxic effects, while ~2% (for 0.33mM) will definitely! If you do short term treatment, i.e. minute to a few hours it may still be OK, though. As controls you would need to but 0.6% and 2% DMSO into separate dishes respectively.
My advice would be to either try to dissolve your drug e.g. in H2O or at a much higher concentration (at least 10-fold) in DMSO.
Hope that helps!
I always make 1000X concentration of the treatment concentration. then I will use 1ul per ml of media correspondingly I will take 1ul of DMSO per ml media. this way it is easier to add for different volume.
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