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Assay Length
4.5 hours
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Sample Type & Volume Required Per Well
Cell Culture Supernates (100 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL)
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Sensitivity
18.3 pg/mL
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Assay Range
31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Specificity
Natural and recombinant human IL-1ra
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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
Interference observed with 1 or more available related molecules.
Control Available
QC25, Quantikine Immunoassay Control Set 810 for Human IL-1ra -
Product Summary
The Quantikine Human IL-1ra Immunoassay is a 4.5 hour solid phase ELISA designed to measure IL-1ra in cell culture supernates, serum, and plasma. It contains E. coli-derived recombinant human IL-1ra as well as antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate recombinant human IL-1ra. Results obtained using natural human IL-1ra showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-1ra.
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 known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean | 64.9 | 538 | 1343 | 56.2 | 469 | 1243 |
| Standard Deviation | 3.67 | 22.1 | 55.7 | 5.79 | 31.9 | 70.4 |
| CV% | 5.6 | 4.1 | 4.1 | 10.3 | 6.8 | 5.7 |
Serum, EDTA Plasma, Heparin Plasma | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean | 66.9 | 607 | 1627 | 82.5 | 646 | 1663 |
| Standard Deviation | 4.88 | 30.2 | 60 | 9.04 | 51.4 | 111 |
| CV% | 7.3 | 5 | 3.7 | 11 | 8 | 6.7 |
Recovery
The recovery of IL-1ra spiked to three levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Supernates (n=4) | 106 | 96-115 |
| EDTA Plasma (n=4) | 86 | 66-102 |
| Heparin Plasma (n=4) | 93 | 82-106 |
| Serum (n=4) | 90 | 81-99 |
Linearity
To assess the linearity of the assay, samples containing high concentrations of IL-1ra were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Background: IL-1ra/IL-1F3
IL-1ra (IL-1 receptor antagonist) is a widely expressed acute phase protein that serves to dampen inflammation. IL-1ra binds to both IL-1 RI and IL-1 RII. It competitively inhibits IL-1 (alpha or beta) binding to IL-1 RI and does not trigger recruitment of the accessory molecule IL-1 R AcP or activation of IL-1 RI signaling. IL-1ra blocks the proinflammatory actions of IL-1 including Prostaglandin E2 and IL-6 release from endothelial cells, fever generation, thrombocytosis, the production of hepatic acute phase proteins, and neutrophil expansion and infiltration.
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