MTT analysis using GraphPad Prism

1. Edit the concentration of the drug/treatment under consideration. NB: Excel sheet formula auto-populates the required parameters.
   

2. Fill the respective absorbance read for each concentration.

3. Copy the concentration (in µg) to the “µg” column. (Here, the concentration will be converted from µg to G. Same formula can be used as concentration used was measured in µM.

4. The LogC(G) column generated will be the X-axis for the analysis and the viabilities (Not their averages) will be the value for the Y-axis.

Working in GraphPad Prism

1. In Prism, select:

   • XY

   • Option > Data Table (Select enter or import data into new table)

   • Under Options > X : Select “Numbers”

   • Under Options > Y : Select “replicate values in a side-by-side sub-columns and enter the number of replicates. ( select Ok after)

2. Copy LogC(G) > axis variable values into X columns

3. Copy variabilities into first group ( Group A) and if there are other groups, copy them into their various groups.

4. Select ” Analyze” in the analysis tab

   • Under XY analysis select:

     • Nonlinear regression (curve fit) > Ok

5. Go to Graphs

   • Click on data and for plot select “Mean and error” , “SEM”.