Species:
Universal
Alternative Name:
ARA
Assay Type:
Competitive
unit:
μg/mL
Standard:
200 μg/mL
Sensitivity:
0.94 μg/mL
Detection Range:
3.13-200 μg/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma, Tissue homogenates, Cell lysates, Cell culture supernates and Other biological fluids
Research:
Signal transduction;Metabolic pathway;Endocrinology;Neuro science;Hormone metabolism;Bone metabolism;
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with AA. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to AA. 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 AA in the samples is then determined by comparing the OD of the samples to the standard curve.
| μg/mL | OD | Corrected |
|---|---|---|
| 200 | 0.174 | |
| 100 | 0.31 | |
| 50 | 0.525 | |
| 25 | 0.859 | |
| 12.5 | 1.287 | |
| 6.25 | 1.679 | |
| 3.13 | 2.055 | |
| 0 | 2.495 |
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) | 3.94 | 8.4 | 56.18 | 4.1 | 6.29 | 69.93 |
| Standard deviation | 0.42 | 0.93 | 2.9 | 0.52 | 1.15 | 3.61 |
| C V (%) | 3.32 | 4.48 | 4.15 | 4.02 | 4.92 | 2.5 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 90-105 | 97 |
| EDTA plasma (n=8) | 88-102 | 95 |
| Cell culture media (n=8) | 87-97 | 90 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 92-105 | 89-98 | 92-103 |
| Average (%) | 97 | 94 | 94 | |
| 1:4 | Range (%) | 92-101 | 89-98 | 86-97 |
| Average (%) | 100 | 94 | 87 | |
| 1:8 | Range (%) | 80-94 | 81-94 | 83-96 |
| Average (%) | 85 | 91 | 85 | |
| 1:16 | Range (%) | 86-101 | 96-105 | 91-103 |
| Average (%) | 89 | 104 | 99 |