Species:
Universal
Alternative Name:
Homocysteine
Assay Type:
Competitive
unit:
ng/mL
Standard:
8000 ng/mL
Sensitivity:
39.55 ng/mL
Detection Range:
125-8000 ng/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma and Other biological fluids
Research:
atherosclerosis
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with HCy. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to HCy. 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 HCy in the samples is then determined by comparing the OD of the samples to the standard curve.
| ng/mL | OD | Corrected |
|---|---|---|
| 8000 | 0.203 | |
| 4000 | 0.36 | |
| 2000 | 0.57 | |
| 1000 | 0.834 | |
| 500 | 1.118 | |
| 250 | 1.418 | |
| 125 | 1.752 | |
| 0 | 2.548 |
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) | 157.48 | 336.13 | 2247.19 | 163.93 | 251.57 | 2797.2 |
| Standard deviation | 16.78 | 37.08 | 116.07 | 20.89 | 46.16 | 144.48 |
| C V (%) | 5.4 | 4.65 | 2.43 | 5.54 | 7.52 | 5.86 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 80-95 | 87 |
| EDTA plasma (n=8) | 87-95 | 91 |
| Cell culture media (n=8) | 82-97 | 89 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 93-102 | 87-98 | 85-97 |
| Average (%) | 97 | 96 | 96 | |
| 1:4 | Range (%) | 93-106 | 87-98 | 90-99 |
| Average (%) | 99 | 87 | 90 | |
| 1:8 | Range (%) | 87-101 | 83-94 | 82-101 |
| Average (%) | 97 | 86 | 94 | |
| 1:16 | Range (%) | 91-104 | 85-98 | 93-105 |
| Average (%) | 99 | 88 | 99 |