Technical Data
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
Concentration |
OD1 |
OD2 |
Average OD |
Corrected |
20 |
2.352 |
2.353 |
2.353 |
2.284 |
10 |
1.558 |
1.564 |
1.561 |
1.49 |
5 |
0.927 |
0.935 |
0.931 |
0.859 |
2.5 |
0.444 |
0.448 |
0.446 |
0.376 |
1.25 |
0.265 |
0.267 |
0.266 |
0.197 |
0.63 |
0.165 |
0.167 |
0.166 |
0.097 |
0.32 |
0.118 |
0.118 |
0.118 |
0.05 |
0 |
0.068 |
0.068 |
0.068 |
0 |
Linear
Samples were spiked with high concentrations of target proteins and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
|
|
Serum (n=5) |
EDTA plasma (n=5) |
Cell culture media (n=5) |
1:2 |
Range (%) |
85-92 |
95-103 |
85-94 |
Average (%) |
90 |
97 |
88 |
1:4 |
Range (%) |
83-96 |
80-92 |
97-106 |
Average (%) |
88 |
81 |
99 |
1:8 |
Range (%) |
80-98 |
97-105 |
82-95 |
Average (%) |
84 |
102 |
85 |
1:16 |
Range (%) |
90-99 |
89-97 |
93-105 |
Average (%) |
91 |
90 |
94 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level were tested on 3 different plates, 20 replicates in each plate, respectively.
Item |
Intra-assay Precision |
Intra-assay Precision |
Sample |
3 |
3 |
Replicate |
9 |
18 |
CV(%) |
5 |
8 |
Rate of recovery
The recovery of spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
Sample Type |
Range(%) |
Average Recovery(%) |
Serum (n=8) |
82-95 |
88 |
EDTA plasma (n=8) |
89-103 |
96 |
Cell culture media (n=8) |
80-93 |
85 |