| Product Name | Rat IL5(Interleukin 5) ELISA Kit |
| Alternative Names | IL-5;E-CSF; ESGF; EDF; TRF; Colony-Stimulating Factor,Eosinophil; T-Cell replacing Factor; B Cell Differentiation Factor I; Eosinophil Differentiation Factor |
| Standard | 500 pg/mL |
| UniProt ID | Q08125 |
| Sample Type | serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids |
| Assay Type | Sandwich |
| Research Area | Cytokine; Infection immunity |
| Assay length | 3.5h |
Test Principle
The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Rat IL5. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat IL5. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Rat IL5, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. 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 Rat IL5 in the samples is then determined by comparing the OD of the samples to the standard curve.
Standard Curve
| Concentration (pg/mL) | OD | Corrected OD |
| 500.00 | 2.156 | 2.067 |
| 250.00 | 1.755 | 1.666 |
| 125.00 | 1.198 | 1.109 |
| 62.50 | 0.797 | 0.708 |
| 31.25 | 0.543 | 0.454 |
| 15.63 | 0.313 | 0.224 |
| 7.82 | 0.196 | 0.107 |
| 0.00 | 0.089 | 0.000 |
Precision
- Intra-assay Precision (Precision within an assay)：CV%<8%
- Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
- Inter-assay Precision (Precision between assays)：CV%<10%
- Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.
Recovery
Matrices listed below were spiked with certain level of recombinant IL5 and the recovery rates were calculated by comparing the measured value to the expected amount of IL5 in samples.
| Matrix | Recovery Range(%) | Average Recovery(%) |
| serum(n=5) | 85-97% | 91% |
| EDTA plasma(n=5) | 83-99% | 95% |
| Heparin plasma(n=5) | 85-97% | 91% |
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of IL5 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
| Matrix | 1:2 | 1:4 | 1:8 | 1:16 |
| serum(n=5) | 88-104% | 84-98% | 95-105% | 88-102% |
| EDTA plasma(n=5) | 78-92% | 83-96% | 95-107% | 87-102% |
| Heparin plasma(n=5) | 87-98% | 92-105% | 95-107% | 82-96% |
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