Hello Elmira,

As you have already got the absorbance by ELISA in MTT analysis, put the values on MS Excel sheet as you have calculated IC50 values.

First, get the average of triplicates of every single dose (drug1+drug2)

Calculate the Cytotoxicity by dividing the Average by the Control value.

Get the % Cytotoxicity, multiplying with 100.

Otherwise, you can get the Cell Viability = (Control-Average)/Control

And get The % Cell viability as well.

When you get all the percentiles compared with Control values put them in Graphpad prism for getting the graph of % cytotoxicity and % of cell viability

At which point %cytotoxicity matches with %cell viability, that's your IC50 of synergistic effect.

The process will be similar in the case of a single and more than one drug.

Thank you,

Dear Elmira!

Please You follow the link

https://www.graphpad.com/support/faq/how-can-i-figure-out-if-two-drugs-are-additive-or-synergistic/

https://www.graphpad.com/support/faq/how-can-i-figure-out-if-two-drugs-are-additive-or-synergistic/

Probably this link may help you

Dear Elmira Zarei,

You can use SynergyFinder online tool to perform such analysis:

https://synergyfinder.fimm.fi/synergy/20230119134555694642/