The compound used for MTT is not soluble in water and sparingly soluble in 5% DMSO, due to which there is issue with uptake in the cells. How can this be solved?
if you are taking about MTT salt, then we generally dissolve in PBS. If it is your compound of interest den try preparing higher concentration of ur compound in 100%DMSO or methanol. Use DMSO as control in MTT assay
Hello Mansi,
Some compounds are not soluble in water and sparingly soluble in DMSO. You can also try dissolving your compound in ethanol as most of the compounds do dissolve in ehtanol. But try to run a paralled control of cells with maximum volume of solvent used per well. Name of the compound will help more in this context.
So first observe that in which solvent, your compound is dissolving maximum. Once you sort out this and still a significant amount remain undissolved then incubating the compound at 37 for 1 hour or overnight can help to increase solubility. If the compound is still not dissolving then look for the permisable amounts of the concentration to be used of that compound/ml of the solvent. Sometimes we use high conc. which are not recommended and hence our solution gets saturated and compound don't dissolve beyond that point.
Hope it helps !
MTT and solubility are two different things. For compounds one can try water, DMSO or ethanol. Try to dissolve your compound in 100% DMSO or Ethanol. Check maximum ethanol conc. that cells will tolerate and prepare dilutions accordingly. don't forget to keep solvent control. Otherwise, from the stock prepare dilutions in medium used for cells provided there is no precipitation of compound in the medium. Hope this will help you.
Dear all, thanks for your kind reply.
I have tried dissolving my test compound in 100% DMSO, 100% ethanol and even in chloroform, but I am not able to make it completely soluble. This is posing a major problem while giving treatment to cells as the compound is settling down in the wells and no uptake is seen,
Please suggest me how can I troubleshoot solubility.
Hello Mansi,
Sometimes single system don't work well in complete dissolution of a compound. You can try using 70% ethanol and 30% DMSO and then dissolve your compound in this mixture. Don't forget to incubate at 37 for sometime.
Hope it helps !
As discussed earlier, I guess, the concentration of your preparation is higher than the maximum solubility of the tetrazolium salt. In our lab we use to prepare 5 mg/ml solution in PBS or in a tissue culture media lacking phenol red and never had any issue with solubility.
Dissolving in solvents like DMSO or EtOH will increase the solubility. However, it will have an issue with cell viability as both DMSO and EtOH are lethal for cells at certain concentrations. You have to optimize the concentrations of these solvents before the treatment.
Hi, 1X PBS is better to dissolve MTT which we use regularly and getting good results. And also, preparing it freshly before the time adding is quite considerable. Good Luck
Guys Comeon.....She is not asking about dissolution of MTT............
eVERYBODY KNOWS IT DISSOLVES EASILY..sHE IS ASKING ABOUT solubility of some compound whose MTT assay she wants to perform...
Wake up...why you people are making answers without understanding the question properly ???????
cHEERS !
Yes my concern was regarding the solubility of my test compound for MTT assay.
It's OK Ajay. Challenging of cell line with some component and MTT assay are two different things. The question was not clear about that. So there is no issue about answering without understanding????????
However,
@ Mansi: May I know the name of component and the concentration at which you wants to dissolve it.
Hi Namita, I am treating cells with a metal complex and I have tried range of solvents for that including 1-5% DMSO, 20%ethanol, acetone and even in cell culture media but everytime it setlles down and once I filter it, the compound is lost.
Can you suggest me any alternative for such difficult to dissolve compounds whose intrinsic property I need to check inside cells?
Thanks
Hi Namita, I am treating cells with a metal complex and I have tried range of solvents for that including 1-5% DMSO, 20%ethanol, acetone and even in cell culture media but everytime it setlles down and once I filter it, the compound is lost.
Can you suggest me any alternative for such difficult to dissolve compounds whose intrinsic property I need to check inside cells?
Thanks
Well I don't have experience with metals. Choice of the salt and pH may effect the solubility.
not sure if this helps you, but there are some links on RG about somewhat similar problem.
https://www.researchgate.net/post/How_do_you_dissolve_ZnSO4_heptahydrate_in_cell_culture_medium
https://www.researchgate.net/post/Are_there_any_good_pH_7_5-8_buffer_that_does_not_form_precipitate_with_metals2
https://www.researchgate.net/post/How_can_I_dissolve_ZnCl2_in_water
Good Luck
@ Namita Rokana,
Yes dear, there is an issue deifnitely. We can't address a problem properly if we don't understand it. It will only result into piling up of unimportant things without any substantial solution. There will be loss of time also and other people will carry on answering based on the same assumption as previous fellow has answered (same has happened here too). Please take it positively...
Thanks !
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