Dear all:
I'd like to treat our MSC with deferoxamine mesylate, DFO (Sigma), so that HIF-a is stablized. After reading its infomation sheet, I'm still clue-less on how to handle it.
I'm planning to prepare deferoxamine mesylate (DFO) at 100mM, as 1000X stock in water, and add 1 microliter to each 1 milileter culture medium. I'll bring DFO to room temperature 1 hour prior to opening it.
According to its infomation sheet, DFO will "decompose on exposure to air", suggesting me dissolve all DFO at once. However with my schedule of experiments, only a small amount of DFO stock is needed at at time, yet accoring to info sheet again, "solutions deteriorate on storage and should be prepared immediately prior to use" tells me not to use up DFO at once.
Would you kindly share your experience with me, please? I really really appreciate it!
It's been long, and no one else answered, please excuse me for talking to myself.
I ended up dissolving DFO in ddH2O, and put it through 0.22um syringe filter to make it sterile.
The 100mM stock was aliquoted in black micro tubes and sealed with parafilm, frozen in -20C frig. To avoid repeated freeze-thaw cycles, I aliquoted it in very small volume, so that I can use it up within 2nd thawing. In this way, it worked just fine.
Hope this information could be useful to someone :)
Yes DFO can be dissolved in cold double distilled water. Then it must be frozen in one time use aliquots. to treat your cells I would recommend 100uM of DFO for 24 hours or up to 1000uM. But 1000 might kill them. For cell lines I often use 500uM.
Thank you Yi-chun. it was really useful of sharing the prep of DFO.
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