| Product Name | Rat IL6(Interleukin 6) ELISA Kit |
| Alternative Names | IL-6;MGI2-A; MGI2A; HGF; BSF2; HSF; IFNB2; B-Cell Stimulatory Factor-2; Hybridoma/Plasmacytoma Growth Factor; Hepatocyte Stimulating Factor; Cytotoxic T-Cell Differentiation Factor |
| Standard | 500 pg/mL |
| UniProt ID | P20607 |
| Sample Type | serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids |
| Assay Type | Sandwich |
| Research Area | Cytokine; Tumor immunity; Infection immunity; Cardiovascular biology |
| 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 IL6. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat IL6. 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 IL6, 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 IL6 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 | 2.153 | 2.068 |
| 250.00 | 1.628 | 1.543 |
| 125.00 | 1.243 | 1.158 |
| 62.50 | 0.948 | 0.863 |
| 31.25 | 0.507 | 0.422 |
| 15.63 | 0.357 | 0.272 |
| 7.82 | 0.215 | 0.130 |
| 0.00 | 0.085 | 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 IL6 and the recovery rates were calculated by comparing the measured value to the expected amount of IL6 in samples.
| Matrix | Recovery Range(%) | Average Recovery(%) |
| serum(n=5) | 81-95% | 88% |
| EDTA plasma(n=5) | 87-99% | 93% |
| Heparin plasma(n=5) | 87-99% | 93% |
Linearity
| Matrix | 1:2 | 1:4 | 1:8 | 1:16 |
| serum(n=5) | 89-97% | 85-94% | 79-96% | 85-99% |
| EDTA plasma(n=5) | 86-98% | 91-103% | 85-97% | 87-96% |
| Heparin plasma(n=5) | 87-95% | 85-97% | 89-101% | 85-97% |
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