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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
4.25 hours or 4.5 hours
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Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
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Sensitivity
2 pg/mL
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Assay Range
3.9 - 250 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Specificity
Natural and recombinant human IL-15
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Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
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Interference
No significant interference observed with available related molecules.
Control Available
QC26, Quantikine Immunoassay Control Set 934 for Human IL-15 -
Product Summary
The Quantikine Human IL-15 Immunoassay is a 4.25 or 4.5 hour solid phase ELISA designed to measure human IL-15 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human IL-15 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural human IL-15 expressed by HuT-102 cells 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-15.
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 | 7.2 | 22.9 | 131 | 9.2 | 29 | 154 |
Standard Deviation | 0.3 | 1.3 | 5.8 | 0.7 | 1.6 | 10.2 |
CV% | 4.2 | 5.7 | 4.4 | 7.6 | 5.5 | 6.6 |
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 | 7.7 | 24.7 | 140 | 9.9 | 32.8 | 184 |
Standard Deviation | 0.3 | 1.3 | 4.8 | 0.9 | 2.7 | 12.2 |
CV% | 3.9 | 5.3 | 3.4 | 9.1 | 8.2 | 6.6 |
Recovery
The recovery of human IL-15 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) | 98 | 93-104 |
EDTA Plasma (n=5) | 99 | 90-112 |
Heparin Plasma (n=5) | 100 | 91-107 |
Serum (n=5) | 98 | 92-109 |
Linearity
To assess the linearity of the assay, five samples were spiked with high concentrations of IL-15 in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.\line
Preparation and Storage
Background: IL-15
IL-15 (Interleukin 15) is a cytokine that induces or enhances the differentiation, maintenance, or activation of multiple T cell subsets including NK, NKT, Th17, Treg, and CD8+ memory cells. It also induces dendritic cell differentiation and inflammatory activation, exhibits anti-tumor activity, and inhibits the deposition of lipid in adipocytes. Complexes of IL-15 with cell surface IL-15 R alpha interact with complexes of IL-2 R beta and the Common gamma Chain on adjacent cells. This transpresentation mechanism enables cells to respond to IL-15 even if they do not express IL-15 R alpha. Ligation of membrane-associated IL-15/IL-15 R alpha complexes also induces reverse signaling that promotes activation of the IL-15/IL-15 R alpha expressing cells. The activity of circulating IL-15 is limited by its association with soluble IL-15 R alpha.