Species:
Universal
Alternative Name:
Cysteinyl Leukotrienes E4
Assay Type:
Competitive
unit:
pg/mL
Standard:
2000 pg/mL
Sensitivity:
9.41 pg/mL
Detection Range:
31.25-2000 pg/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma, Tissue homogenates, Cell culture supernates and Other biological fluids
Research:
Metabolic pathway;Infection immunity;Autoimmunity;
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with LTE4. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to LTE4. 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 LTE4 in the samples is then determined by comparing the OD of the samples to the standard curve.
| pg/mL | OD | Corrected |
|---|---|---|
| 2000 | 0.186 | |
| 1000 | 0.322 | |
| 500 | 0.537 | |
| 250 | 0.871 | |
| 125 | 1.299 | |
| 62.5 | 1.691 | |
| 31.25 | 2.067 | |
| 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) | 39.37 | 84.03 | 561.8 | 40.98 | 62.89 | 699.3 |
| Standard deviation | 4.2 | 9.27 | 29.02 | 5.22 | 11.54 | 36.12 |
| C V (%) | 5.44 | 4.38 | 4.53 | 5.36 | 5.39 | 5.12 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 80-92 | 85 |
| EDTA plasma (n=8) | 87-99 | 93 |
| Cell culture media (n=8) | 85-99 | 92 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 88-97 | 86-97 | 96-105 |
| Average (%) | 97 | 87 | 97 | |
| 1:4 | Range (%) | 91-105 | 86-97 | 89-98 |
| Average (%) | 97 | 96 | 95 | |
| 1:8 | Range (%) | 87-98 | 83-96 | 81-94 |
| Average (%) | 97 | 92 | 94 | |
| 1:16 | Range (%) | 95-105 | 91-103 | 96-105 |
| Average (%) | 103 | 97 | 100 |