We want to dissolve our research samples in cell culture media such as demem or emem. We want to conduct bio-assay guided technique to isolate bio-active compounds. But we have a difficulty to dissolve the samples in the media. We also added 1% of DMSO in the media but the samples were not dissolving. I want an assistant from researchers conducting a similar research.
Dear Professor Aman!
If your compounds are water-insoluble, you can dissolve them in pure (100%, 99.96% DMSO) or ethyl alcohol, and then add the required volume to DMEM or EMEM, depending on the medium in which the cells grow.
Hello Aman
If your research samples are soluble in DMSO or any other solvent, make a highly concentrated solution of your samples in that solvent. For instance, If substance A is soluble in DMSO make a 1mg/ml stock solution of substance A in DMSO. Initially try to dissolve substance A in minimum quantity of DMSO as follows
For 1mg/ml stock solution
Dissolve 10 mg of substance A in 1 ml DMSO and make up the volume to 10 ml by adding 9ml culture medium (DMEM or EMEM).
If substance A dissolves in 2ml DMSO make up the volume to 10ml by adding 8ml culture medium and so on and so forth.
Now you will have a stock solution of 1mg/ml of substance A dissolved in minimum quantity of DMSO Use this stock solution in the required quantity (whatever concentration you desire) in your culture medium (DMEM or EMEM) that will finally be used in the bio-assay.
Hope this is helpful.
Thank you.
Adding protein to your medium may improve the solubility of your substances. If you are able to use a small amount of serum you could try adding fetal bovine serum (FBS) to a desired concentration, say 1% - 10% v/v. Otherwise, you could try adding bovine serum albumin (BSA), from a powder, to a desired concentration of say 0.1% to 1%. Sterile filter the medium when all additions have been completed. You may be able to find filtration products from companies such as GIBCO. I have not done this for a while, but I did routinely sterile filter my culture media. The overall process required access to an autoclave to sterilize my filtration apparatus.
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