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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)
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Sensitivity
57.4 pg/mL
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Assay Range
125.0 - 4,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Specificity
Natural and recombinant human IL-13
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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.
Control Available
QC20, Quantikine Immunoassay Control Group 3 -
Product Summary
The Quantikine Human IL-13 Immunoassay is a 4.5 hour solid phase ELISA designed to measure IL-13 levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human IL-13 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate recombinant human IL-13 accurately. Results obtained using natural human IL-13 showed dose response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-13.
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 | 1 | 2 | 2 | 3 | 3 | 1 | 1 | 2 | 2 | 3 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 360 | 287 | 800 | 1031 | 2002 | 1602 | 319 | 297 | 818 | 932 | 1895 | 1662 |
| Standard Deviation | 12.9 | 18 | 39.3 | 27.5 | 43.2 | 63.1 | 22.2 | 33.6 | 79.3 | 52.6 | 108 | 162 |
| CV% | 3.6 | 6.3 | 4.9 | 2.7 | 2.2 | 3.9 | 7 | 11.3 | 9.7 | 5.6 | 5.7 | 9.7 |
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 | 352 | 960 | 1919 | 365 | 1087 | 2199 |
| Standard Deviation | 11.1 | 21.3 | 33.3 | 25.8 | 63.1 | 105 |
| CV% | 3.2 | 2.2 | 1.7 | 7.1 | 5.8 | 4.8 |
Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 335 | 878 | 1737 | 327 | 898 | 1906 |
| Standard Deviation | 21 | 54 | 40.9 | 20.6 | 34 | 124 |
| CV% | 6.3 | 6.2 | 2.4 | 6.3 | 3.8 | 6.5 |
Recovery
The recovery of IL-13 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=4) | 108 | 100-120 |
| EDTA Plasma (n=4) | 106 | 98-120 |
| Heparin Plasma (n=4) | 104 | 94-120 |
| Serum (n=4) | 102 | 95-113 |
Linearity
To assess linearity of the assay, samples were spiked with high concentrations of IL-13 and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Background: IL-13
IL-13 (Interleukin-13) plays a key role in the pathogenesis of allergy, cancer, and tissue fibrosis. It is secreted by several helper T cell subsets, NK cells, mast cells, eosinophils, basophils, and visceral smooth muscle cells. IL-13 suppresses the production of proinflammatory cytokines and other cytotoxic substances by macrophages, fibroblasts, and endothelial cells. It promotes B cell activation, immunoglobulin class switching to IgE, and the upregulation of CD23/Fc epsilon RII. IL-13 signals through a receptor complex containing IL-13 R alpha 1 and IL-4 R alpha. This complex also functions as the type 2 IL-4 receptor. IL-13 R alpha 2 binds IL-13 and prevents IL-13 from signaling through the IL-13 R alpha 1/IL-4 R alpha complex. IL-13-bound IL-13 R alpha 2 can directly promote tumor cell invasiveness and the development of tissue fibrosis.
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