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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), Urine (100 uL)
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Sensitivity
1.08 pg/mL
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Assay Range
3.9 - 250 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine)
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Specificity
Natural and recombinant human IL-5
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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
QC20, Quantikine Immunoassay Control Group 3 -
Product Summary
The Quantikine Human IL-5 Immunoassay is a 4.5 hour solid phase ELISA designed to measure IL-5 levels in cell culture supernates, serum, plasma, and urine. It contains Sf 21-expressed recombinant human IL-5 and antibodies raised against the recombinant factor and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human IL-5 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 IL-5.
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.
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 | 31.3 | 95.6 | 173 | 32.9 | 97.9 | 180 |
Standard Deviation | 1.4 | 3.5 | 6.5 | 4 | 6.7 | 8.4 |
CV% | 4.5 | 3.7 | 3.8 | 12.2 | 6.8 | 4.7 |
Cell Culture Supernates, Urine | Intra-Assay Precision | Inter-Assay Precision |
Sample | 1 | 2 | 3 | 1 | 2 | 3 |
n | 20 | 20 | 20 | 40 | 40 | 40 |
Mean | 28.1 | 89.3 | 170 | 31.2 | 92.6 | 174 |
Standard Deviation | 0.89 | 2 | 7.27 | 2.5 | 4.2 | 8.7 |
CV% | 3.2 | 2.2 | 4.2 | 8 | 4.5 | 5 |
Recovery
The recovery of human IL-5 spiked to different levels throughout the range of the assay in various matrices was evaluated.
Sample Type | Average % Recovery | Range % |
Cell Culture Media (n=4) | 101 | 95-107 |
EDTA Plasma (n=4) | 104 | 86-118 |
Heparin Plasma (n=4) | 101 | 87-112 |
Serum (n=4) | 104 | 88-116 |
Urine (n=4) | 96 | 91-106 |
Linearity
To assess the linearity of the assay, five samples were spiked with high concentrations of IL-5 in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Background: IL-5
IL-5 (Interleukin-5) is a secreted cytokine that is expressed as a covalent dimer. It is produced by activated Th2 cells, eosinophils, mast cells, and iNKT cells and promotes eosinophilic and basophilic differentiation and activation. IL-5 signals through a receptor complex containing IL-5 R alpha and the Common beta Chain/CD131 on eosinophils and basophils. Naturally occurring soluble forms of IL-5 R alpha retain the ability to bind IL-5 and limit its bioavailability.