Species:
Universal
Alternative Name:
F-E3; Free Estriol
Assay Type:
Competitive
unit:
ng/mL
Standard:
40 ng/mL
Sensitivity:
0.218 ng/mL
Detection Range:
0.63-40 ng/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma, Tissue homogenates, Cell lysates, Cell culture supernates and Other biological fluids
Research:
Endocrinology;Reproductive science;Hormone metabolism;
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with F-E3. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to F-E3. 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 F-E3 in the samples is then determined by comparing the OD of the samples to the standard curve.
| ng/mL | OD | Corrected |
|---|---|---|
| 40 | 0.258 | |
| 20 | 0.421 | |
| 10 | 0.661 | |
| 5 | 0.978 | |
| 2.5 | 1.271 | |
| 1.25 | 1.591 | |
| 0.63 | 1.861 | |
| 0 | 2.683 |
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) | 0.79 | 1.68 | 11.24 | 0.82 | 1.26 | 13.99 |
| Standard deviation | 0.08 | 0.19 | 0.58 | 0.1 | 0.23 | 0.72 |
| C V (%) | 3.67 | 6.74 | 4.1 | 3.33 | 5.61 | 4.31 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 90-105 | 97 |
| EDTA plasma (n=8) | 80-95 | 87 |
| Cell culture media (n=8) | 83-95 | 89 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 80-91 | 86-97 | 92-105 |
| Average (%) | 81 | 97 | 93 | |
| 1:4 | Range (%) | 89-103 | 86-97 | 87-98 |
| Average (%) | 90 | 88 | 95 | |
| 1:8 | Range (%) | 82-95 | 90-98 | 85-92 |
| Average (%) | 92 | 97 | 91 | |
| 1:16 | Range (%) | 86-99 | 80-93 | 80-96 |
| Average (%) | 94 | 87 | 95 |