Bio-Rad Experion Protein Analysis Kits - Manual (Page 20)

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3. Examine the separation of at least one sample. Click on the sample name in the project tree or on
the lane in the virtual gel to view the electropherogram, which should have the following features:
n
Two well-resolved marker peaks and a set of system peaks
n
Sample peaks located between the system peaks and upper marker
n
Flat baseline
If the upper and lower marker peaks are not properly assigned, you may need to include or exclude
peaks, or manually set the marker (see Section 6.3).
4. Evaluate the data analysis performed by Experion software (see Chapter 4, Data Analysis). If
necessary, change the analysis settings and parameters — including protein quantitation methods
— by following the instructions in Chapters 5 and 6.
3.10 Evaluate the Run
When a run is complete, evaluate the run and the analysis of the data by Experion software.
1. Ensure all lanes (ladder and samples) are visible in the virtual gel. The markers (indicated by pink
triangles) should be visible in and aligned across all lanes. If the marker peaks are not properly
assigned, you may need to include or exclude peaks, or manually set the marker (see Section 6.3).
2. Evaluate the separation of the Pro260 ladder. To display the ladder electropherogram, click the
ladder well in the project tree, or click on the lane labeled L in the virtual gel. The electropherogram
should resemble the one shown in Figure 3.6 and should have the following features (if your ladder
does not have these features, see Chapter 7, Troubleshooting for more information):
n
Two marker peaks and a set of system peaks
n
Eight Pro260 ladder peaks between the system peaks and upper marker
n
Flat baseline
n
Marker peaks at least 20 fluorescence units above the baseline
Experion Automated Electrophoresis System
Fig . 3 .6 . Separation of the Pro260 ladder . Note
the flat baseline and well-resolved peaks. All
identified peaks are numbered, and the lower and
upper markers are indicated by green asterisks (*).
The 260 kD protein in the Pro260 ladder is labeled
as the upper marker in the Results table.
Pro260 ladder peaks
Baseline
Upper marker
Lower marker
System peaks