Species:
Universal
Alternative Name:
LT-B4
Assay Type:
Competitive
unit:
pg/mL
Standard:
1000 pg/mL
Sensitivity:
4.39 pg/mL
Detection Range:
15.63-1000 pg/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma, Tissue homogenates, Cell culture supernates and Other biological fluids
Research:
Metabolic pathway;Infection immunity;Endocrinology;Hormone metabolism;
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with LTB4. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to LTB4. 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 LTB4 in the samples is then determined by comparing the OD of the samples to the standard curve.
| pg/mL | OD | Corrected |
|---|---|---|
| 1000 | 0.188 | |
| 500 | 0.324 | |
| 250 | 0.539 | |
| 125 | 0.873 | |
| 62.5 | 1.301 | |
| 31.25 | 1.693 | |
| 15.63 | 2.069 | |
| 0 | 2.509 |
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) | 393.7 | 840.34 | 5617.98 | 409.84 | 628.93 | 6993.01 |
| Standard deviation | 41.95 | 92.71 | 290.18 | 52.22 | 115.4 | 361.21 |
| C V (%) | 4.02 | 4.57 | 5.4 | 5.96 | 7 | 4.6 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 89-101 | 95 |
| EDTA plasma (n=8) | 82-95 | 88 |
| Cell culture media (n=8) | 80-94 | 87 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 87-98 | 93-102 | 82-95 |
| Average (%) | 92 | 96 | 86 | |
| 1:4 | Range (%) | 92-101 | 93-102 | 88-103 |
| Average (%) | 96 | 94 | 92 | |
| 1:8 | Range (%) | 82-98 | 81-96 | 86-97 |
| Average (%) | 92 | 82 | 93 | |
| 1:16 | Range (%) | 89-97 | 93-106 | 87-102 |
| Average (%) | 94 | 97 | 92 |