I am doing an MTT Assay of a new drug that I am experimenting with on MDAMB-231 Cells; however, I cannot figure out a dilution factor or an experimental setup on the MTT assay to see the IC50 in-between a 25 micromolar and a 12.5 micromolar concentration. The reason I am testing this is I ran an MTT assay before and it showed that the IC50 is between 25 micromolar and 12.5. So how can I set up MTT so that different stock solutions DO NOT have to be made for each column? Please let me know as I have until this Friday 2/18/2022 to let my PI know.
I would suggest preparing your drug at a lower stock concentration (e.g. 1.25 mM [100X for 12.5 µM]). You can then add the drug to separate tubes of media to generate the intermediate concentrations that should cover your IC50. Using a 1.25 mM concentration will allow you to easily make 9 different concentrations between 12.5 µM and 25 µM using increments of 1 µl.
Start at 200uM and do 3 1:2 dilutions down to 25uM. Then do 3 3:4 dilutions down to 10.5uM, then finish up with 3 more 1:2 dilutions.
Kristin M. Adams, I like your idea, however, the cell line I am working with can not exceed a DMSO percentage of 0.3% as I am working with MDAMB231 cells.
John Hardy Lockhart, I am having trouble visualizing your idea. Can you draw it out for me? I would appreciate it.
Kristin M. Adams How do you do a 3:4 dilution from a serial dilution while marinating the same volume in each well? Please let me know thank you.
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