Species:
Universal
Alternative Name:
PTD
Assay Type:
Competitive
unit:
ng/mL
Standard:
400 ng/mL
Sensitivity:
2.13 ng/mL
Detection Range:
6.25-400 ng/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma and Other biological fluids
Research:
Metabolic pathway;Endocrinology;
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with PTD. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to PTD. 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 PTD in the samples is then determined by comparing the OD of the samples to the standard curve.
| ng/mL | OD | Corrected |
|---|---|---|
| 400 | 0.241 | |
| 200 | 0.404 | |
| 100 | 0.644 | |
| 50 | 0.961 | |
| 25 | 1.254 | |
| 12.5 | 1.574 | |
| 6.25 | 1.844 | |
| 0 | 2.666 |
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) | 7.87 | 16.81 | 112.36 | 8.2 | 12.58 | 139.86 |
| Standard deviation | 0.84 | 1.85 | 5.8 | 1.04 | 2.31 | 7.22 |
| C V (%) | 4.93 | 5.77 | 4.42 | 4.62 | 7.53 | 2.93 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 88-99 | 93 |
| EDTA plasma (n=8) | 86-98 | 92 |
| Cell culture media (n=8) | 82-98 | 87 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 83-94 | 85-93 | 87-99 |
| Average (%) | 90 | 91 | 95 | |
| 1:4 | Range (%) | 80-92 | 85-93 | 82-96 |
| Average (%) | 81 | 86 | 86 | |
| 1:8 | Range (%) | 86-102 | 80-96 | 85-96 |
| Average (%) | 96 | 90 | 91 | |
| 1:16 | Range (%) | 87-103 | 93-106 | 87-101 |
| Average (%) | 87 | 103 | 90 |