Hi! I will be using Amphotericin B as a positive control for disc diffusion assay (DDA) towards C. albicans, in SD Agar.
I need to inoculate 25µg/20μL of Amphotericin B in each disc.
I aim to use 0.1% DMSO to dissolve Amphotericin B powder.
Will this work?
How do I dilute the 25 mg Amphotericin B powder to 25µg/20μL solution?
Thanks for the help in advance!
Hi Jonathan,
according to Sigma instructions, you can dissolve amphotericin B in water at pH 2 or 11. However, consider that this pH are extreme for most organisms. You should prepare using a buffer with low concentration, so once dissolved, the cell culture media bufers it to neutral pH.
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/134/925/a9528dat.pdf
Also, you can use 100% DMSO to dissolve up to 30 - 40 mg/ml. Again, DMSO could be toxic for cells, so you should not add more than 1/200 (0,5%), depending on the cell type.
Your concentration would be 1,25 ug/uL, or 1,25 mg/ml (25/20). So, I would dissolve in water at pH 2 or 11, as DMSO may kill your cells. Alternatively, use a control of DMSO alone to check that the effect you observe is caused by Amphotericin B, and not the DMSO.
Good luck!
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