This kit is based on Double antibody-Sandwich ELISA detection method and takes 4h assay time. The microplate provided in this kit has been precoated with anti IL-15 antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antibody. Then, it binds with IL-15 bound to precoated antibody. Wash unbound components and add HRP-Streptavidin Conjugate (SABC). Wash unbound components again and add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Calculate the concentration of IL-15 in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.
Specification
| Product Name | Rat IL-15(Interleukin-15) ELISA Kit |
| Alias | Interleukin-15 ELISA Kit, IL-15 ELISA Kit, IL15 ELISA Kit |
| UniProt ID | P9760 |
| Sample Type | Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples |
| Detection Method | Sandwich ELISA, Double Antibody |
| Detection Wavelength | OD450 |
| Reaction Duration | 4 hours |
| Storage | 2-8°C(Sealed), Don't cryopreserve. |
| Specificity | Specifically binds with IL-15 , no obvious cross reaction with other analogues. |
Standard Curve
This product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18°C -25°C; Equilibrate TMB substrate to 37°C before staining. After adding into the ELISA wells, incubate for 15min at 37°C in dark.)
Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.
| STD.(pg/ml) | OD-1 | OD-2 | Average |
| 0 | 0.113 | 0.117 | 0.115 |
| 15.625 | 0.134 | 0.138 | 0.136 |
| 31.25 | 0.176 | 0.182 | 0.179 |
| 62.5 | 0.258 | 0.266 | 0.262 |
| 125 | 0.583 | 0.599 | 0.591 |
| 250 | 0.959 | 0.987 | 0.973 |
| 500 | 1.66 | 1.708 | 1.684 |
| 1000 | 2.402 | 2.472 | 2.437 |
Recovery
Add a certain amount of IL-15 into the sample. Calculate the recovery by comparing the measured value with the expected amount of IL-15 in the sample.
| Sample Type | Recovery Range(%) | Average Recovery(%) |
| serum(n=10) | 86-98 | 91 |
| EDTA plasma(n=10) | 87-98 | 91 |
| Heparin plasma(n=10) | 86-104 | 93 |
Linearity
Dilute the sample with a certain amount of IL-15 at 1:2, 1:4 and 1:8 to get the recovery range.
| Sample Type | 1:2 | 1:4 | 1:8 |
| serum(n=10) | 87-102% | 91-104% | 96-104% |
| EDTA plasma(n=10) | 83-100% | 86-97% | 84-101% |
| Heparin plasma(n=10) | 81-95% | 93-100% | 97-100% |
Precision(%)
Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.
Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.
| Item | Intra-assay Precision | Inter-assay Precision | ||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/ml) | 31.65 | 131.5 | 506 | 32.68 | 127.4 | 506 |
| Standard deviation | 1.66 | 6.88 | 24.39 | 2.07 | 7.9 | 29.9 |
| CV(%) | 5.23 | 5.23 | 4.82 | 6.32 | 6.2 | 5.91 |
Stability
Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.
| ELISA kit(n=5) | 37°C for 1 month | 2-8°C for 6 months |
| Average(%) | 80 | 95-100 |
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