Species:
Universal
Alternative Name:
2-(3,4-Dihydroxyphenyl)ethylamine; 3,4-Dihydroxyphenethylamine; 3-Hydroxytyramine; Intropin; Revivan; Oxytyramine; Dopamine Hydrochloride
Assay Type:
Competitive
unit:
pg/mL
Standard:
1000 pg/mL
Sensitivity:
4.71 pg/mL
Detection Range:
15.63-1000 pg/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma and Other biological fluids
Research:
Signal transduction;Cardiovascular biology;Reproductive science;Neuro science;
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with DA. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to DA. 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 DA in the samples is then determined by comparing the OD of the samples to the standard curve.
| pg/mL | OD | Corrected |
|---|---|---|
| 1000 | 0.284 | |
| 500 | 0.384 | |
| 250 | 0.578 | |
| 125 | 0.839 | |
| 62.5 | 1.134 | |
| 31.25 | 1.529 | |
| 15.63 | 1.843 | |
| 0 | 2.507 |
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) | 19.69 | 42.02 | 280.9 | 20.49 | 31.45 | 349.65 |
| Standard deviation | 2.1 | 4.64 | 14.51 | 2.61 | 5.77 | 18.06 |
| C V (%) | 3.4 | 4.59 | 3.24 | 3.83 | 5.59 | 5.35 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 80-95 | 87 |
| EDTA plasma (n=8) | 86-99 | 92 |
| Cell culture media (n=8) | 80-95 | 87 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 88-97 | 84-99 | 93-101 |
| Average (%) | 96 | 94 | 101 | |
| 1:4 | Range (%) | 91-105 | 84-99 | 89-105 |
| Average (%) | 96 | 95 | 89 | |
| 1:8 | Range (%) | 87-98 | 83-101 | 87-98 |
| Average (%) | 94 | 86 | 93 | |
| 1:16 | Range (%) | 95-105 | 86-99 | 92-100 |
| Average (%) | 99 | 95 | 100 |