Bio-Rad Experion Protein Analysis Kits - Manual (Page 30)

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
£
10010510A.qxp 2/13/2008 11:43 AM Page 26