7. Quality Control
108
Instructions for Use for INFINITE M1000 PRO No. 30064852 Rev. No. 1.0
2011-09
7.4.3
Fluorescence Polarization (FP)
7.4.3.1 FP Precision Fluorescein 384-well Plate:
Pipette 100 µl of the reagents into the wells of a Greiner 384-well plate (black, flat
bottom) according to Plate Layout.
Material:
Greiner 384-well plate, flat bottom, black
100 µl pipette (8-channel) + tips
1 nM Fluorescein
0.01 nM NaOH (=Blank)
High Polarization Standard (Invitrogen)
Plate Layout:
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1
2
3
4
5
6
7
8
9
10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
A
F P F B F B F B F B F B
B
C
F P F B F B F B F B F B
D
E
F B F B F B F B F B F B
F
G
F B F B F B F B F B F B
H
I
F B F B F B F B F B F B
J
K
F B F B F B F B F B F B
L
M
F B F B F B F B F B F B
N
O
F B F B F B F B F B F B
P
(F….100 µl 1 nM Fluorescein; B…..100 µl 0.01 M NaOH = Blank, P…100 µl
Invitrogen High Pol Standard)
Measurement Parameters:
Measurement mode:
Fluorescence Polarization
Excitation wavelength:
470 nm
Emission wavelength:
525 nm
Emission bandwidth:
20 nm
Gain:
‘Optimal’
Lag
time: 0
µs
Number of reads:
10 (= 100 ms integration time)
Settle
time:
50
ms
Plate definition file:
GRE384fb.pdfx
Z-Position:
Calculated from Well A1
G-Factor reference:
Calibrated from well A1, ref. value = 20 mP;
G-Factor reference blank:
I3; sample blank same as reference blank
Evaluation:
Calculate the lower detection limit [mP] = 'precision':
FP Precision (mP) = Stdev
F
Stdev
F
Standard deviation of wells filled with Fluorescein (except reference well)
