Bio-Rad Experion Protein Analysis Kits - Manual (Page 37)

4. After evaluating the run as described in Section 3.10, select Analysis > Internal Std . and Std
Protein Calibration Curve.
5. In the Internal Std . and Std . Protein Calibration Curve window (Figure 5.5), do the following:
a. Under Standard Information, next to Standard Protein Molecular Weight, enter the MW
corresponding to the region where the software searches for a peak, for both the calibrants
and the samples. The software checks for a peak in the sample wells that has the entered
value (± 5%). If necessary, modify the concentrations of the calibrants by double-clicking on the
concentration value and typing in a new value.
b. Ensure that Use User Internal Standard is not selected.
c. Under Calibration Curve Settings, if the protein used for calibration (calibrant) is different or
has a different mass than the protein being quantitated, select Quantitate All Proteins and click
Apply. If you do not do this, the software only reports absolute concentrations for peaks that are
the same size as the calibrant.
d. Under Calibration Curve Settings, select the Linear Regression to be used.
e. Click Apply to update the calculation. The calibration curve appears and displays the equation
for the linear regression (Figure 5.5). The table below the graphic displays data for the wells
containing the standards. Click Close.
6. In the Results tab, the concentration for each protein identified by Experion software is derived
by relative quantitation against the 260 kD upper marker and automatically appears in the
Concentration column; the calibrated concentrations of proteins found in the specified region
appear in the Calib . Conc . column.
Technical Support: 1-800-4BIORAD • 1-800-424-6723 • www.bio-rad.com 33
Fig . 5 .5 . Internal Std . and Std Protein Calibration Curve window . In this example, a 100 kD protein is used to build a
5-point calibration curve for quantitating a 100 kD sample protein.
Experion Pro260 Analysis Kit