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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 (50 uL), Serum (50 uL)
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Sensitivity
7 pg/mL
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Assay Range
15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum)
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Specificity
Natural and recombinant mouse 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.
Product Summary
The Quantikine Mouse IL-5 Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse IL-5 in cell culture supernates and mouse serum. It contains Sf 21-expressed recombinant mouse IL-5 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate recombinant mouse IL-5. Results obtained using natural mouse IL-5 showed linear curves that were parallel to the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse 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.
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 | 41.5 | 117 | 428 | 43.4 | 123 | 447 |
Standard Deviation | 2.4 | 3.9 | 8.6 | 1.8 | 5.3 | 20.8 |
CV% | 5.8 | 3.3 | 2 | 4.1 | 4.3 | 4.7 |
Recovery
The recovery of mouse IL-5 spiked to three levels throughout the range of the assay in various matrices was evaluated.
Sample Type | Average % Recovery | Range % |
Cell Culture Supernates (n=5) | 99 | 85-110 |
Serum (n=5) | 92 | 81-104 |
Linearity
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of mouse IL-5 in each matrix were diluted with Calibrator Diluent and then assayed.
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.