54
Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516
TM397 · Revised 12/16
www.promega.com
3.E. Analysis of Results (continued)
4.
Performing an Analysis (continued)
h. Performing the analysis will generate a curve fit from the data on the graphing tab of the ‘Analysis’ screen.
The graphing tab will remain inactive until the
Run
button has been pressed. Depending on the type of
curve fit being applied to the data, the following information will be created as a result of pressing the
Run
button.
Linear Standard Curve
The average value of the blanks will be subtracted from all known and unknown samples on the plate map.
Averages and Standard Deviations (if there are greater than three replicates of each concentration) will be
calculated for standard curve replicates, and a linear fit (y = mx + b) of the standards will be generated on
the graphing tab.
Slope, Y-intercept, and r
2
values for the linear fit of the standard curve replicates are calculated and
displayed on the graphing tab.
The linear curve fit equation is used to interpolate the values of each of the unknowns on the plate.
Multiplying the interpolated concentration by the dilution factor for the unknown will generate the
interpolated concentration for each unknown well. Concentrations are displayed for each unknown well.
Averages and standard deviations (assuming there are three or more replicates) will be calculated for each
unknown replicate set and will be indicated in the export file for the analysis.
Linear Response
The average value of the blanks will be subtracted from all known samples on the plate map.
Averages and Standard Deviations (if there are greater than 3 replicates of each concentration) will be
calculated for each replicate concentration set, and a linear fit (y = mx + b) of the experimental data set will
be generated on the graphing tab.
Slope, Y-intercept, and r
2
values for the linear fit of the experimental data set replicates are calculated and
displayed on the graphing tab.
Averages and standard deviations (assuming there are 3 or more replicates) for each sample replicate set
will be indicated in the export file for the analysis.
Dose Response–EC
50
/IC
50
The average value of the blanks will be subtracted from all knowns and unknowns on the plate map.
Averages and Standard Deviations (if there are greater than 3 replicates of each concentration) will be
calculated for the experimental data set replicates, and a four-parameter curve fit
(y = B + (T – B/1 + 10
[logx] + s
) of the experimental data set will be generated on the graphing tab.
EC
50
/IC
50
, Top, Bottom and Slope values for the four-parameter fit of the experimental data set replicates
are calculated and displayed on the graphing tab.
Averages and standard deviations (assuming there are 3 or more replicates) for each replicate set will be
indicated in the export file for the analysis.
