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Assay Type
Solid Phase Sandwich ELISA
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Format
96-well strip plate
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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), Citrate Plasma (100 uL)
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Sensitivity
15 pg/mL
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Assay Range
31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
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Specificity
Natural and recombinant human IL-17
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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.
Control Available
QC21, Quantikine Immunoassay Control Group 4 -
Product Summary
The Quantikine Human IL-17 Immunoassay is a 4.5 hour solid phase ELISA designed to measure human IL-17 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human IL-17 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate recombinant human IL-17 accurately. Results obtained using natural human IL-17 showed linear 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 human IL-17.
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 | 20 | 20 | 20 |
| Mean | 107 | 604 | 1217 | 76.6 | 272 | 549 |
| Standard Deviation | 7.8 | 25 | 63.3 | 6.5 | 23.4 | 43.9 |
| CV% | 7.3 | 4.1 | 5.2 | 8.5 | 8.6 | 8 |
Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 131 | 736 | 1480 | 125 | 408 | 794 |
| Standard Deviation | 5.8 | 34.6 | 61.2 | 9.8 | 34.3 | 55.6 |
| CV% | 4.4 | 4.7 | 4.1 | 7.8 | 8.4 | 7 |
Recovery
The recovery of human IL-17 spiked to three different levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Media (n=5) | 97 | 88-112 |
| Citrate Plasma (n=5) | 105 | 92-116 |
| EDTA Plasma (n=5) | 101 | 90-110 |
| Heparin Plasma (n=5) | 101 | 93-109 |
| Serum (n=5) | 98 | 90-105 |
Linearity
To assess the linearity of the assay, five samples spiked with high concentrations of IL-17 were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Background: IL-17/IL-17A
The IL-17 family is comprised of at least six proinflammatory cytokines that share a conserved cysteine-knot structure but diverge at the N-terminus. IL-17 family members are glycoproteins secreted as dimers that induce local cytokine production and recruit granulocytes to sites of inflammation. IL-17 is induced by IL-15 and IL-23, mainly in activated CD4+ T cells distinct from Th1 or Th2 cells. IL-17F is the most homologous to IL-17, but is induced only by IL-23 in activated monocytes. IL-17B binds the IL-17B receptor, but not the IL-17 receptor; it is most homologous with IL-17D, which is expressed by resting CD4+ T cells and CD19+ B cells. IL-17E is mainly produced by Th2 cells and recruits eosinophils to lung tissue. IL-17C has a very restricted expression pattern but has been detected in adult prostate and fetal kidney libraries.
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