Bio-Rad Experion Protein Analysis Kits User Manual
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8.
In the Results table, the concentration for each protein identified by Experion software
is derived by relative quantitation against the 260 kD upper marker and appears in the
Concentration column; the calibrated concentrations of proteins found in the size
region specified appear in the Calib. Conc. column. The expected concentration for
peak 4 (BGG) is 400 ng/µl. In the example in Figure 3.5, relative quantitation against
the upper marker yielded a concentration of 574.0 ng/µl, while absolute quantitation
yielded a concentration of 413.1 ng/µl.
Fig. 3.5. Results table from Test 3 showing relative quantitation (Concentration) and absolute
quantitation (Calib. Conc.) calculations.
9.
To evaluate the reproducibility of absolute quantitation, determine the %CV for wells
1–3, 9, and 10. Copy the data to Excel software and determine %CV as the standard
deviation/mean x 100. Values should be £20%.
10. Calculate the mean concentration determined for the unknown samples. Note that the
absolute quantitation method generates values that are more accurate (closer to the
expected value of 400 ng/µl) than those derived by relative quantitation. Accuracy
should be within 20% for 400 ng/µl BGG.
Expected Performance of BGG for Test 3
Measurement
Typical Results
Linear regression (calibration curve)
r
2
0.97
Absolute quantitation
Reproducibility
%CV £ 20%
Accuracy
400 ± 80 ng/µl
26
£
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