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| Product Name | Chicken IgY(Immunoglobulin Y) ELISA Kit | Species | Chicken |
| Uniprot ID | N/A | Alternative Names | |
| Detection method | Sandwich | Sensitivity | 0.47 ng/mL |
| Standard | 50ng/mL | Detection Range | 0.78-50ng/mL |
| Sample type | Serum, Plasma, Tissue homogenate and Other biological samples；Sample Volume=100μL | ||
| Reaction time | 3.5H | Research Area | Signal transduction;CD & Adhesion molecule;Apoptosis;Tumor immunity;Infection immunity;Immune molecule; |
| Test principle | This kit uses the Sandwich-ELISA principle. The microtiter plate strips has been pre-coated with an affinity purified antibody to Chicken IgY. Standards or samples containing Chicken IgY are added to the plate and reacted with capture antibody. A second anti-Chicken IgY antibody labeled biotin is then added and binds to Chicken IgY captured on the plate. After that, Streptavidin-Horseradish Peroxidase(SA-HRP) is added to form a sandwich complex of solid phase antibody-Chicken IgY-biotin labeled antibody-SA-HRP. And then, TMB substrate solution is added to all wells and incubated. An enzyme-catalyzed reaction generates a blue color in the solution, thereafter, stop solution is added to stop the substrate reaction and the color turns yellow. The yellow solution is read at a wavelength of 450nm. The concentration of Chicken IgY in the samples is then calculated from the OD value by establishing a standard curve. | ||
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| (ng/mL) | OD | Corrected |
|---|---|---|
| 50.00 | 2.512 | 2.435 |
| 25.00 | 1.713 | 1.579 |
| 12.50 | 1.013 | 0.913 |
| 6.25 | 0.464 | 0.394 |
| 3.13 | 0.228 | 0.185 |
| 1.57 | 0.132 | 0.087 |
| 0.79 | 0.085 | 0.035 |
| 0.00 | 0.052 |
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) | 3.05 | 12.25 | 22.32 | 3.26 | 12.56 | 22.79 |
| Standard deviation | 0.15 | 0.61 | 1.12 | 0.16 | 0.63 | 1.14 |
| C V (%) | 4.75 | 2.66 | 6.05 | 4.81 | 2.73 | 6.23 |
The recovery of spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 92-106 | 99 |
| EDTA plasma (n=8) | 88-103 | 95 |
| Cell culture media (n=8) | 80-97 | 88 |
Samples were spiked with high concentrations of target proteins and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 83-97 | 87-96 | 81-90 |
| Average (%) | 89 | 88 | 90 | |
| 1:4 | Range (%) | 82-96 | 87-96 | 82-96 |
| Average (%) | 92 | 93 | 95 | |
| 1:8 | Range (%) | 86-98 | 86-95 | 85-98 |
| Average (%) | 97 | 91 | 96 | |
| 1:16 | Range (%) | 93-101 | 92-103 | 92-103 |
| Average (%) | 100 | 102 | 93 |