Species:
Universal
Alternative Name:
FTI
Assay Type:
Competitive
unit:
pg/mL
Standard:
90 pg/mL
Sensitivity:
0.48 pg/mL
Detection Range:
1.41-90 pg/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma, Tissue homogenates, Cell lysates, Cell culture supernates and Other biological fluids
Research:
Signal transduction;
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with fT4. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to fT4. Next,Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of fT4 in the samples is then determined by comparing the OD of the samples to the standard curve.
| pg/mL | OD | Corrected |
|---|---|---|
| 90 | 0.444 | |
| 45 | 0.56 | |
| 22.5 | 0.756 | |
| 11.25 | 1.041 | |
| 5.63 | 1.379 | |
| 2.81 | 1.694 | |
| 1.41 | 1.93 | |
| 0 | 2.449 |
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.
| Intra-assay Precision | Inter-assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean(ng/mL) | 1.77 | 3.78 | 25.28 | 1.84 | 2.83 | 31.47 |
| Standard deviation | 0.19 | 0.42 | 1.31 | 0.23 | 0.52 | 1.63 |
| C V (%) | 6.51 | 5.49 | 4.65 | 5.93 | 4.66 | 2.23 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 87-99 | 93 |
| EDTA plasma (n=8) | 80-93 | 85 |
| Cell culture media (n=8) | 85-97 | 91 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 88-104 | 85-92 | 90-99 |
| Average (%) | 104 | 91 | 97 | |
| 1:4 | Range (%) | 93-102 | 85-92 | 95-103 |
| Average (%) | 101 | 90 | 101 | |
| 1:8 | Range (%) | 82-97 | 80-96 | 86-92 |
| Average (%) | 91 | 81 | 86 | |
| 1:16 | Range (%) | 92-101 | 93-105 | 91-102 |
| Average (%) | 95 | 94 | 98 |