According to my paper I have to dissolve carvacrol and thymol in DMSO. Particularly, it says that the substances are dissolved in DMSO and diluted in culture medium. The final concentration of DMSO in cells is 0.1%.
I don't know how to start. How much of DMSO and substance do I have to use ?
You can start from the total amount of DMSO that you need. If the final concentration of DMSO has to be 0.1%, that means that, for example, you will need 100µl DMSO (0.1ml) in 100ml di medium. To be precise, you will have to take 99.9ml medium + 0.1ml DMSO.
Now, if you want to do a stock solution of your two compounds in DMSO, you can think for example to use a concentration 1000x higher than what do you need in the final medium. In this way, adding DMSO at 0.1% (1/1000 of the final volume) you will add the compounds at the right concentration.
Eg: Your carvacrol must be at a concentration of 10ng/ml in the final medium. That means that in 100ml of medium you will nead 1000ng of carvacrol. If you dilute your carvacrol in DMSO at 10*1000=10.000ng/ml , and you need the DMSO at 0.1%, you will need 0.1ml of DMSO. Then, 10.000ng/ml(concentration in DMSO)*0.1ml (volume of DMSO that you use)=1000ng(amount of carvacrol that you needed).
Hope it helped
You can start from the total amount of DMSO that you need. If the final concentration of DMSO has to be 0.1%, that means that, for example, you will need 100µl DMSO (0.1ml) in 100ml di medium. To be precise, you will have to take 99.9ml medium + 0.1ml DMSO.
Now, if you want to do a stock solution of your two compounds in DMSO, you can think for example to use a concentration 1000x higher than what do you need in the final medium. In this way, adding DMSO at 0.1% (1/1000 of the final volume) you will add the compounds at the right concentration.
Eg: Your carvacrol must be at a concentration of 10ng/ml in the final medium. That means that in 100ml of medium you will nead 1000ng of carvacrol. If you dilute your carvacrol in DMSO at 10*1000=10.000ng/ml , and you need the DMSO at 0.1%, you will need 0.1ml of DMSO. Then, 10.000ng/ml(concentration in DMSO)*0.1ml (volume of DMSO that you use)=1000ng(amount of carvacrol that you needed).
Hope it helped
Just a reminder: Dilutions of the stock solution (in DMSO) of the chemical should be prepared freshly prior to performing the experiments.
Also, tested chemicals free wells (negative controls) would require to have the same concentration of medium/DMSO so that you can assure any change is attributable to DMSO interference.
Best regards
Thank you very much for the useful advice and i appeciate the time you spent. According to my paper i have to add 20μL of carvacrol (dissolved in DMSO and diluted in culture medium), 150μl PBMC and after incubation 30μL of LPS. So, in every well the total volume will be 200μL. Do i get the concentration of DMSO for the 200μL volume?
If you are adding 20 uL of carvacrol in DMSO, that would be 20 uL of DMSO without carvacrol you will have to add to negative controls. Am I right?
Does the paper mention the concentration of carvacrol used? It should be in mili, micro or picograms per uL (mg, ug or pg/ul). The less DMSO you add to culture the better, that is why carvacrol dilution in DMSO should have a high concentration (at least 1000 X).
Hi every body
have you ever seen sediment of wogonin-DMSO solution with water?
How about if I want to calculate with known molecular weight? I need to prepare 10 mM stock, and the total volume should be 500 uL. Molecular weight of the chemical is 229.30.
Afroja Rahman : In this case u need to dissolve 1.14 mg of chemical to prepare 10mM stock in 500 ul. For ur ease rearrange formula of molarity for mass of solvent calculation in mg,
mg of solvent=Molarity * Molecular weight* Volume in ul/106 (due to microlitre conversion)
mg of solvent=10 *229.3*500/106
Good Day everyone
How about if I want to prepare urolithin A compounds dissolved in dimethyl sulfoxide (DMSO) at 20 mg/ml and the final concentration of DMSO should not exceed 0.1% in the culture medium. Different concentrations of the compounds, ranging from 15 to 90 𝜇 mol/L are to be used to treat the cell lines.
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