Species:
Universal
Alternative Name:
SM
Assay Type:
Competitive
unit:
μg/mL
Standard:
400 μg/mL
Sensitivity:
1.85 μg/mL
Detection Range:
6.25-400 μg/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma and Other biological fluids
Research:
Signal transduction;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 SPH. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to SPH. 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 SPH in the samples is then determined by comparing the OD of the samples to the standard curve.
| μg/mL | OD | Corrected |
|---|---|---|
| 400 | 0.254 | |
| 200 | 0.417 | |
| 100 | 0.657 | |
| 50 | 0.974 | |
| 25 | 1.267 | |
| 12.5 | 1.587 | |
| 6.25 | 1.857 | |
| 0 | 2.679 |
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 (%) | 3.86 | 5.74 | 3.96 | 5.02 | 7.07 | 4.99 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 80-93 | 85 |
| EDTA plasma (n=8) | 87-99 | 93 |
| Cell culture media (n=8) | 90-103 | 96 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 87-97 | 80-85 | 89-103 |
| Average (%) | 89 | 83 | 92 | |
| 1:4 | Range (%) | 86-92 | 80-85 | 95-102 |
| Average (%) | 90 | 80 | 99 | |
| 1:8 | Range (%) | 89-103 | 87-96 | 96-104 |
| Average (%) | 92 | 92 | 103 | |
| 1:16 | Range (%) | 89-98 | 82-95 | 87-98 |
| Average (%) | 91 | 90 | 88 |