recently I did experiment in culture cell treated with Curcumin. I made a stock of Curcumin with concentration 10 mM from 36 mg of Curcumin powder in 10 ml of DMSO. i treated my cell with concentration of Curcumin 50 mikroMolar from fresh stock of Curcumin 10 mM that I just made, and I got a clear medium in my cell. After that I keep my Curcumin stock at -20 C. today, i'm repeat my experiment and using same doses and curcumin stock that i used before, but in my medium i see black dot ?!(like if your curcumin not dissolve properly). Any suggestion what might be the problem? thank you
There are some questions regarding curcumin solubility here, you could find the answer: https://www.scbt.com/scbt/product/curcumin-458-37-7
We have been using 10 mM curcumin stock prepared in DMSO stored at -20 C without any problems. It is necessary to store the Curcumin solution in dark as it is light sensitive.
I suggest you to check first, the stability of your curcumin stock. This can be done by by measuring its absorption spectra. You can prepare known concentration of curcumin in DMSO and record its absorption and check if there is any change in the absorption maxima. Curcumin is also sensitive to pH changes; you can measure the pH of the cell culture media. If culture media is alkaline, when you add curcumin it under goes colour change.
Dear Queen.
Can be solved through good practice by making fresh CM stock solution (or any plant extract solution to be tested for cells in vitro), centrifuging at 12,000 rpm for 10 min to remove any particles and aggregates and even microscopic strings from the E-tube, taking clear upper supernatant, aliquots in small E-tubes and storing at -70C under aluminium foil.
Thaw completely and take a known vol of CM stock sol, transfer into a known vol of medium containing FBS (i.e., 50ul CM into 450ul medium, 10x dilution, this would be 1st dilution factor), mix well, centrifuge as above, take a known vol of upper sol, (add the vol to known vol of culture medium in a plate well (this would be 2nd dilution factor) to make a final concentration to be tested.
Never put the thawed stock sol back into the freezer. This causes aggregated particle and salting out. There are so many microscopic particle in crude extract solution you made since insoluble compounds are always present. Repeated freezing and thawing enhance the particulates.
So, once you thawed the tube aliquots, throw it away.
You won't need any worries.
Best wishes.
Above points are very useful. I will add that purity of DMSO matters. Its best to use HPLC grade or cell culture grade DMSO for making this stock solution.
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