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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), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Citrate Plasma (50 uL)
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Sensitivity
10 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-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
No significant interference observed with available related molecules.
Control Available
QC01-1, Quantikine Immunoassay Control Group 1 -
Product Summary
The Quantikine Human IL-4 Immunoassay is a 4.5 hour solid phase ELISA designed to measure IL-4 levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human IL-4 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant human IL-4 accurately. Results obtained using natural human IL-4 showed dose-response 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-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 | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean | 159 | 538 | 1092 | 169 | 503 | 1032 |
| Standard Deviation | 11.4 | 27.8 | 67.2 | 16.8 | 49.3 | 83 |
| CV% | 7.2 | 5.2 | 6.2 | 9.9 | 9.8 | 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 | 40 | 40 | 40 |
| Mean | 170 | 510 | 1110 | 132 | 411 | 865 |
| Standard Deviation | 6.7 | 17.5 | 58.5 | 9.8 | 32.1 | 52.8 |
| CV% | 3.9 | 3.4 | 5.3 | 7.4 | 7.8 | 6.1 |
Recovery
The recovery of human IL-4 spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Media (n=8) | 98 | 90-109 |
| Citrate Plasma (n=10) | 94 | 85-109 |
| EDTA Plasma (n=10) | 100 | 90-123 |
| Heparin Plasma (n=10) | 95 | 84-121 |
| Serum (n=10) | 98 | 82-111 |
Linearity
To assess the linearity of the assay, the following biological samples, spiked with high concentrations of IL-4, were diluted with the appropriate 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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