Species:
Universal
Alternative Name:
β-Alanyl-L-Histidine
Assay Type:
Competitive
unit:
ng/mL
Standard:
900 ng/mL
Sensitivity:
3.53 ng/mL
Detection Range:
14.07-900 ng/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma, Tissue homogenates, Cell lysates, Cell culture supernates and Other biological fluids
Research:
Metabolic pathway;Endocrinology;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 Car. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to Car. 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 Car in the samples is then determined by comparing the OD of the samples to the standard curve.
| ng/mL | OD | Corrected |
|---|---|---|
| 900 | 0.422 | |
| 450 | 0.538 | |
| 225 | 0.734 | |
| 112.5 | 1.019 | |
| 56.25 | 1.357 | |
| 28.13 | 1.672 | |
| 14.06 | 1.908 | |
| 0 | 2.427 |
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) | 17.72 | 37.82 | 252.81 | 18.44 | 28.3 | 314.69 |
| Standard deviation | 1.89 | 4.17 | 13.06 | 2.35 | 5.19 | 16.25 |
| C V (%) | 3.87 | 5.46 | 5.69 | 6.71 | 6.6 | 2.46 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 83-95 | 89 |
| EDTA plasma (n=8) | 95-107 | 101 |
| Cell culture media (n=8) | 82-94 | 88 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 87-96 | 87-98 | 89-102 |
| Average (%) | 89 | 89 | 99 | |
| 1:4 | Range (%) | 82-95 | 87-98 | 82-97 |
| Average (%) | 91 | 91 | 85 | |
| 1:8 | Range (%) | 92-105 | 92-101 | 88-105 |
| Average (%) | 94 | 96 | 89 | |
| 1:16 | Range (%) | 89-97 | 82-98 | 83-96 |
| Average (%) | 94 | 83 | 90 |