Dear Community,
as i want to find out the impact of ceramide on my cells in DMEM-F12 medium. I dissolved C16-CER in Ethanol and added it to my medium. After having a look through the microscope, i see that it precipitates.
Does anyone have a solution for my problem?
And do you know if C2-CER is easier to use?
Thanks,
Konstantin
Hi!
Thank you for your answer. I'll try to heat up the medium more than 37 °C before use, but i wonder if precipitation won't appear when it cools down, as i have to maintain my medium at 37 °C during cultivation.
Concentration problem: I used different concentrations from 20 nM to 50 µM. Same picture at every concentration.
BSA solution: i will definitely try to bind C16-CER to BSA, but i'm not very confident that it works, as other publications never mention it.
Could also try a dmso to bsa complex, or dmso. Just to mind to have a control of same except blank with just solvent alone ( etoh, or dmso) warmed.
If you vortex well too with temp, bsa will foam too much for vortex, sonication sounds good, bath.. or probe...
Just as you did, if you see crystals on your monolayer... it wont work well, you want them in solution.
Oh, Also pH may help to mind as well, but your medium will change with the phenol red if it goes beond the 'good' buffer range if its in your dmem.
As i remember most of those sphingolipids are not super soluable in water based solutions. So take your time.
Best wishes, and luck!
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