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Section11:Results
From Assay Guidance Wiki
|
Primary capture antibody |
|||||||||
|---|---|---|---|---|---|---|---|---|---|
|
Secondary Antibody |
2 µg/ml |
1 µg/ml |
0.5 µg/ml |
||||||
|
1:1000 |
H |
L |
0 |
H |
L |
0 |
H |
L |
0 |
|
H |
L |
0 |
H |
L |
0 |
H |
L |
0 |
|
|
1:5000 |
H |
L |
0 |
H |
L |
0 |
H |
L |
0 |
|
H |
L |
0 |
H |
L |
0 |
H |
L |
0 |
|
|
1:25000 |
H |
L |
0 |
H |
L |
0 |
H |
L |
0 |
|
H |
L |
0 |
H |
L |
0 |
H |
L |
0 |
|
Actual results from initial experiment to determine the coating antibody and detection antibody
|
Primary capture antibody |
|||||||||
|---|---|---|---|---|---|---|---|---|---|
|
Secondary Antibody |
2 µg/ml |
1 µg/ml |
0.5 µg/ml |
||||||
|
1:1000 |
3 |
1 |
1 |
3 |
1 |
1 |
3 |
1 |
1 |
|
4 |
1 |
1 |
4 |
1 |
1 |
3 |
1 |
1 |
|
|
1:5000 |
3 |
0.7 |
0.5 |
3 |
0.8 |
0.5 |
3 |
0.8 |
0.4 |
|
3 |
0.7 |
0.5 |
3 |
0.8 |
0.5 |
3 |
0.8 |
0.4 |
|
|
1:25000 |
1 |
0.2 |
0.1 |
1 |
0.2 |
0.1 |
1 |
0.3 |
0.1 |
|
1 |
0.2 |
0.1 |
1 |
0.3 |
0.1 |
2 |
0.3 |
0.1 |
|
Values below are averages at an A.U. of A490.
|
Secondary Antibody Conc. |
50 |
1 |
0 |
50 |
1 |
0 |
50 |
1 |
0 |
|---|---|---|---|---|---|---|---|---|---|
|
1:1000 |
3.69 |
1.81 |
1.37 |
3.63 |
1.89 |
1.33 |
3.3 |
1.79 |
0.99 |
|
1:5000 |
3.22 |
0.7 |
0.49 |
3.24 |
0.81 |
0.47 |
3.1 |
0.83 |
0.36 |
|
1:25000 |
1.61 |
0.21 |
0.15 |
1.75 |
0.25 |
0.16 |
1.72 |
0.26 |
0.12 |
|
Primary Ab ug/ml |
2 |
2 |
2 |
1 |
1 |
1 |
0.5 |
0.5 |
0.5 |
Conclusions: Lowest NSB and best signal to noise ratio from low to high LP276 concentration are the 0.5 µg/ml concentration for the coating antibody and the 1:25,000 dilution of the biotinylated antibody as the detection antibody.
