Dear all,
is there anyone who ever used glutamate to induced SH-SY5Y cell death? I'd like to know the concentration of the glutamate and how to dissolve the glutamate. I learn form literature that the work concentration is from 5mM-100mM, I'd like to make a 50M storage concentration, however, I cannot dissolve it in water. could anyone tell me how to dissolve the glutamate?
thank you!
glutamate is a polar compound and it is quite soluble in water. If not, I propose making a stock solution in DMSO. Then dissolve a small amount of the stock solution in a relatively high amount of culture media to reach your desired concentration. Be careful that the DMSO do not exceed more than 0.1% in your culture media.
50M ?! You can't have that. That would be more concentrated than the pure solid. Try 1M, if you use sodium glutamate.
If you have free glutamic acid, then the developing acidity is going to limit solubility. You'll need to titrate it into solution with NaOH to approximate neutral pH.
Dear all, thanks very much for your answers. what I do now is use medium make 100mM glutamate and then use NaOH titrate it to neutral PH. it works very well.
Instead of diluting Glutamate directtly in the medium, you can first dilute it in higher concentrations in Milli-Q water, adjust the pH if necessary and sterilize it by microfiltration with a 0.20 µm diameter membrane.
For my own experiments, I was making 400 mM stock solution in Milli-Q water, then it was which can be stored in a freezer at -20°C for a month and remains stable.
Afterwards, for your experiments, you can dilute this stock solution to desired concentrations using now a culture medium.
This worked very well for me.
For the concentration to apply to cells, as you said, the range reported in the literature is very wide. Many factors may influence this effect: the study design, the king of medium you use (the effect is not the same for example in MEM or in DMEM), ... Maybe you have already found the appropriate concentration for your experiments, but if not, I could advise you to try different concentrations in this range and see which one matches your needs.
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