L-glutamic acid prep - (Nov/29/2009 )
Hi guys
Anyone out there has prepared L-glutamic acid to add into a neuronal culture to induce excitoxicity before? The glutamic acid i have is from Sigma G8415 and someone told me to dissolve it in H2O and then adjust to pH7.4. However there appears to be some particles left and i had a hard time filtering it thru the 0.2uM filter. I had made it to a 0.3M conc. I tried using it on my neuronal culture at 1mM conc and left it on overnight but it did not cause any noticeable change in the culture compared to the controls. I am wondering if i prepared it wrongly.. I have heard that you are supposed to dissolve it in 1M HCl. Has anyone tried this before? and any idea whether i need to pH it afterwards? Thanks in advance.
cppoo1 on Nov 29 2009, 01:54 PM said:
Anyone out there has prepared L-glutamic acid to add into a neuronal culture to induce excitoxicity before? The glutamic acid i have is from Sigma G8415 and someone told me to dissolve it in H2O and then adjust to pH7.4. However there appears to be some particles left and i had a hard time filtering it thru the 0.2uM filter. I had made it to a 0.3M conc. I tried using it on my neuronal culture at 1mM conc and left it on overnight but it did not cause any noticeable change in the culture compared to the controls. I am wondering if i prepared it wrongly.. I have heard that you are supposed to dissolve it in 1M HCl. Has anyone tried this before? and any idea whether i need to pH it afterwards? Thanks in advance.
Hope this helps
Thanks for the quick reply... it does explain the solubility bit of the glutamic acid but any idea what is the general consensus for its preparation in excitoxicity experiments in neuronal cultures? A lot of papers don't usually explain in detail how they prepare their glutamate..Sigma's website says to dissolve in 1M HCl but i am not very sure if that will be too good for the neurons (in terms of acidity).
Hi I also face the same problem.Any suggestions plz