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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
2 pg/mL
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Assay Range
7.8 - 500 pg/mL (Cell Culture Supernates, Serum)
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Specificity
Natural and recombinant mouse IL-4
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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.
Product Summary
The Quantikine Mouse IL-4 Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse IL-4 levels in cell culture supernates and serum. It contains E. coli-expressed recombinant mouse IL-4 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse IL-4 accurately. Results obtained using natural mouse IL-4 showed dose 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 mouse IL-4.
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 | 20.1 | 58.6 | 199 | 20.7 | 58.8 | 203 |
| Standard Deviation | 1.3 | 1.8 | 8.9 | 1.2 | 2.7 | 10.6 |
| CV% | 6.5 | 3.1 | 4.5 | 5.8 | 4.6 | 5.2 |
Recovery
The recovery of mouse IL-4 spiked to three levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Supernates (n=8) | 95 | 84-109 |
| Serum (n=7) | 95 | 88-118 |
Linearity
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of mouse IL-4 in each matrix were diluted with Calibrator Diluent and then assayed.
Preparation and Storage
Background: IL-4
IL-4 (Interleukin-4) is a cytokine that is primarily expressed by Th2-biased CD4+ T cells, mast cells, basophils, and eosinophils. It promotes cell proliferation, survival, and immunoglobulin class switch to IgG4 and IgE in human B cells, acquisition of the Th2 phenotype by naïve CD4+ T cells, priming and chemotaxis of mast cells, eosinophils, and basophils, and the proliferation and activation of epithelial cells. IL-4 plays a dominant role in the development of allergic inflammation and asthma. The type I receptor for IL-4, which is expressed on hematopoietic cells, is a heterodimer of the ligand binding IL-4 R? and the common ? chain (a shared subunit of the receptors for IL-2, -7, -9, -15, and -21). The type II receptor on nonhematopoietic cells consists of IL-4 R? and IL-13 R?1. The type II receptor also transduces IL-13 mediated signals.
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