Species:
Universal
Alternative Name:
Histamin
Assay Type:
Competitive
unit:
ng/mL
Standard:
100 ng/mL
Sensitivity:
0.51 ng/mL
Detection Range:
1.57-100 ng/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma, Tissue homogenates, Cell lysates, Cell culture supernates and Other biological fluids
Research:
Signal transduction;Infection immunity;
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with HA. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to HA. 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 HA in the samples is then determined by comparing the OD of the samples to the standard curve.
| ng/mL | OD | Corrected |
|---|---|---|
| 100 | 0.268 | |
| 50 | 0.368 | |
| 25 | 0.562 | |
| 12.5 | 0.823 | |
| 6.25 | 1.118 | |
| 3.13 | 1.513 | |
| 1.56 | 1.827 | |
| 0 | 2.491 |
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) | 1.97 | 4.2 | 28.09 | 2.05 | 3.14 | 34.97 |
| Standard deviation | 0.21 | 0.46 | 1.45 | 0.26 | 0.58 | 1.81 |
| C V (%) | 6.67 | 7.35 | 5.23 | 4.63 | 5.53 | 5.49 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 90-105 | 97 |
| EDTA plasma (n=8) | 80-95 | 87 |
| Cell culture media (n=8) | 88-102 | 95 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 81-96 | 90-99 | 91-99 |
| Average (%) | 85 | 98 | 96 | |
| 1:4 | Range (%) | 86-93 | 90-99 | 89-97 |
| Average (%) | 89 | 94 | 94 | |
| 1:8 | Range (%) | 82-95 | 87-103 | 95-105 |
| Average (%) | 94 | 98 | 100 | |
| 1:16 | Range (%) | 80-94 | 83-96 | 87-102 |
| Average (%) | 90 | 93 | 100 |