Hi,
I ran an MTT assay with 7 doses of my drug ( dissolved in DMSO , final concentration in media ,0.05%) , and read the absorbance at 490nm according to the reagent manufacturers instructions. one problem that i have is that when i want to plot my concentrations to the absorbance my 0% control has an absorbance between two of my doses. my drug exerts a proliferative effect on neural stem cells and increases their proliferation by approximately 2.5 fold. yet the last two high concentrations of the drug has a negative effect and kills the cells. i don't have a 100% control ( i couldn't find an equivalent molecule for my drug). one more thing i have a dose of 1 micromolar and when i want to transform my data to its logarithmic form, i can't choose 1 for both my 0 control and my specified dose. what should i do? can i use these data to plot an EC50 graph??
and one more thing would you guys please describe me how to best design an assay for determining EC50 of a drug? thanks alot.
Dear Neda, here is one of possibly useful answers: https://www.researchgate.net/post/How_to_choose_concentration_range_of_drug_for_cancer-cell_proliferation_assay
Also, there are some suggestions regarding calculations in Excel: https://www.researchgate.net/post/Can_someone_help_out_on_how_i_can_use_excel_to_determine_ic50_in_cytotoxic_cell_growth
thanks dear. Vilma. that was sorta helpful. but my problem is more concerned with incorporating my blank control results in the data obtained from MTT assay. my blank control ( no drug ) or in some online references refered to as 100% control has an absorbance between doses 10 and 15 micromolar and not maximum result. thus it totally messes up my graph. can i exclude my blank control (Cell with DMSO 0.05% but no drug) and still draw an EC50 graph? because i did try it and the graph looks fine. but is it scientifically accurate to draw a graph with its blank control?
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