1 protein quantitation methods, 2 performing percentage determination – Bio-Rad Experion Protein Analysis Kits User Manual
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5.1 Protein Quantitation Methods
Experion
™
software enables protein quantitation by several approaches. The selection of approaches
depends on the type of experiment and desired level of accuracy (Table 5.1). Refer to Appendix A and
bulletin 5784 for details about the different quantitation methods.
Table 5.1. Protein quantitation methods used by Experion software. Absolute quantitation methods provide greater
accuracy than relative quantitation methods. The accuracy of percentage determination is protein dependent.
Calibration Method and Internal Standard Used
Single-Point
Calibration Curve
Quantitation Method
Output
Upper Marker
User-Defined
Upper Marker
User-Defined
Percentage Determination
% Total
No internal standard required
Concentration Determination
Relative quantitation
ng/µl
l
—
—
—
—
l
—
—
Absolute quantitation
ng/µl
—
—
l
—
—
—
—
l
5.2 Performing Percentage Determination
Experion software automatically calculates a percentage of total protein (% total) value for each detected
peak and lists the results in the % Total column of the Results table (Figure 5.1). Click on the peak of
interest in the electropherogram to highlight the data for that peak.
Fig . 5 .1 . Results table showing quantitation data for peaks in a sample . Experion software automatically
performs % total and relative concentration determinations for each peak identified in every sample.
To exclude a peak from the % total calculation, right-click on the peak (in either the single-well
electropherogram view or Results table) and select Exclude Peak in the context menu. The new
calculations for the remaining peaks appear in the Results table.
This change affects a selected sample. To exclude multiple peaks from multiple samples, use the
Settings tab and the Peak Find Settings options in the All Wells Settings dialog (see Section
6.4, Excluding a Peak From Analysis and 6.5, Changing Peak Finding Parameters) and adjust the
peak detection criteria.
Experion Automated Electrophoresis System