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Assay Length
4.5 hours
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Sample Type & Volume Required Per Well
Cell Culture Supernates (25 uL), Serum (25 uL), EDTA Plasma (25 uL), Heparin Plasma (25 uL)
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Sensitivity
5.22 pg/mL
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Assay Range
15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Specificity
Natural and recombinant mouse IL-10
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Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
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Interference
No significant interference observed with available related molecules.
Product Summary
The Quantikine Mouse IL-10 Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse IL-10 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse IL-10 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse IL-10 accurately. Results obtained using natural mouse IL-10 showed dose response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse IL-10.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of a known concentration were tested in separate assays to assess inter-assay precision over two lots of kit components. Assays were performed by at least three technicians.
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 76.3 | 123 | 815 | 71.4 | 115 | 809 |
| Standard Deviation | 2.56 | 2.85 | 17 | 5.39 | 7.64 | 55.5 |
| CV% | 3.4 | 2.3 | 2.1 | 7.5 | 6.6 | 6.9 |
Cell Culture Supernates, Serum | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 26.8 | 75.5 | 585 | 26.2 | 80 | 583 |
| Standard Deviation | 2.5 | 6 | 29 | 1.7 | 6.2 | 31 |
| CV% | 9.3 | 7.9 | 5 | 6.5 | 7.8 | 5.3 |
Recovery
The recovery of IL-10 spiked to levels throughout the range of the assay was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Samples (n=4) | 99 | 95-103 |
| EDTA Plasma (n=4) | 91 | 86-99 |
| Heparin Plasma (n=4) | 89 | 82-98 |
| Serum (n=4) | 89 | 85-96 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with various concentrations of mouse IL-10 in each matrix were diluted with Calibrator Diluent and then assayed.
Preparation and Storage
Background: IL-10
Interleukin-10 functions as an anti-inflammatory cytokine by inhibiting the expansion and activation of Th1 cells and Th17 cells and by promoting the development of M2 macrophages and regulatory T cells (Treg). Within a tumor microrenvironment, however, IL-10 can inhibit the expansion of both Treg and myeloid-derived suppressor cells (MDSC). IL-10 exerts protective effects including limiting tissue damage in arthritic inflammation and promoting muscle regeneration after injury, but it also contributes to the persistence of viral infections. IL-10 signals through a receptor complex composed of IL-10 R alpha and IL-10 R beta. IL-10 R beta additionally associates with IL-20 R alpha, IL-22 R alpha 1, or IL-28 R alpha to form the receptor complexes for IL-22, IL-26, IL-28, and IL-29.
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