7. Quality Control
114
Instructions for Use for INFINITE M1000 PRO No. 30064852 Rev. No. 1.0
2011-09
Evaluation:
Calculate the detection limit in fmol/well:
15
B
ATP
B
8
1e
1
*
0.00001
*
mean
mean
Stdev
*
3
*
10
2
well)
imit(fmol/
DetectionL
−
−
−
⋅
=
2*10
-8
Concentration of ATP standard [M]
Stdev
B
Standard deviation of Blank (B: A3 – D9)
mean
ATP
Average of wells filled with ATP standard
mean
B
Average of Blank wells (B: A3 – D9)
0.00001
Conversion into mol/well
1/1e
-15
Conversion into fmol/well
7.4.4.5 Crosstalk ATP 1536-well Plate:
Material:
Same plate/plate layout and reagent as described in chapter 7.4.4.4 Detection
Limit ATP 1536-well Plate
Evaluation:
Calculate Crosstalk from measurement results of chapter 7.4.4.4 Detection Limit
ATP 1536-well Plate for each well separately. The following wells are charged
together: A2 with A1, A2 with A3, B2 with B1, B2 with B3, C2 with C1, C2 with
C3, D2 with D1, and D2 with D3. The average of these 8 values must be within
the specified limits, see chapter 7.3 Acceptance Criteria (under Luminescence
Crosstalk).
)
(
100
*
)
(
%
B
ATP
B
B
CT
wellx
X
−
−
=
B
X
Blank wells A1 – D1 and A3 – D3, respectively
ATP
wellx
ATP wells A2 – D2
B
Average of wells A4- D10 (Blank)
7.4.5 AlphaScreen
7.4.5.1 AlphaScreen Detection Limit:
Pipette the reagents into the wells of a Greiner 384-well plate (white, flat bottom)
according to the Plate Layout
Material:
AlphaScreen Omnibeads #6760626D (PerkinElmer)
Greiner 384-well plate, flat bottom, white
Phosphate-buffered saline (PBS)
10 µl p tips
100 µl p tips
Omnibeads dilution series:
Dilute the Omnibeads stock solution 1:500 in PBS by adding 3 µl of the stock
solution (5 mg/ml) to 1497 µl PBS (yielding a solution of 10 µg/ml).
Prepare 12 further dilutions in 1:2 steps by pipetting 750 µl of the previous
dilution step to 750 µl PBS. Use a new tip for each dilution step.