7.
Quality
Control
2011-09
Instructions for Use for INFINITE M1000 PRO No. 30064852 Rev. No. 1.0
115
Plate layout:
Pipette 100 µl of each dilution into 5 replicate wells of the microplate (as shown in
the Plate Layout). Use 100 µl PBS for the Blank wells.
Use a fresh tip for each concentration and take care NOT to contaminate
the blank with any Omnibeads dilution!
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A
10 µg/ml
B
5 µg/ml
C
2.5 µg/ml
D
1.25 µg/ml
E
0.62 µg/ml
F
0.31 µg/ml
G
0.15 µg/ml
H
0.08 µg/ml
I
0.04 µg/ml
J
0.02 µg/ml
K
0.01 µg/ml
L
0.005 µg/ml
M
0.002 µg/ml
N
Blank (PBS only)
O
P
Filling volume: 100 µl/well of each Omnibeads dilution (5 replicate wells each) or
blank (PBS only)
Measurement Parameters:
Before pipetting the plate, prepare instrument for measurement:
Parameters:
Measurement mode:
AlphaScreen/AlphaLISA
Excitation time:
100 ms
Integration time:
300 ms
Temperature correction: activated
Plate definition file:
GRE384fw.pdfx
Part of the plate:
A2 – P6
Start measurement immediately after pipetting!
Evaluation:
Calculate the average and standard deviation for each Omnibeads concentration.
Perform a blank reduction by subtracting the average signal of the blanks wells
from the average signal of each Omnibeads concentration.
Plot the average blank-corrected values against the final Omnibeads
concentrations in a XY scatter diagram. Add a linear trend line with intercept set
to 0 and solve the trend line equation (
y = kx
) using the 3-fold standard deviation
of the blank as y.
k
y
x
=
y
3* stdev of the blank (wells N2-P6)
Extrapolate the detection limit [ng/ml] by using the 3-fold standard deviation of the
blank as y.
