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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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Specificity
Natural and recombinant rat IL-2
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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 Rat IL-2 Immunoassay is a 4.5 hour solid phase ELISA designed to measure rat IL-2 levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant rat IL-2 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural rat IL-2 showed dose response 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 rat IL-2.
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, 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 | 130 | 478 | 1491 | 139 | 495 | 1550 |
| Standard Deviation | 4.2 | 10.5 | 31.9 | 13.8 | 30.8 | 85.4 |
| CV% | 3.2 | 2.2 | 2.1 | 9.9 | 6.2 | 5.5 |
Recovery
The recovery of rat IL-2 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) | 110 | 93-120 |
| EDTA Plasma (n=4) | 102 | 90-117 |
| Heparin Plasma (n=4) | 100 | 92-116 |
| Serum (n=14) | 96 | 83-114 |
Linearity
To assess the linearity of the assay, four or more samples containing and/or spiked with various concentrations of rat IL-2 were diluted with Calibrator Diluent and then assayed.
Preparation and Storage
Background: IL-2
IL-2 (Interleukin 2) is a critical cytokine in T cell biology. It promotes T cell activation and expansion, the development, maintenance and function of regulatory T cells (Treg), and the differentiation of CD8+ T cells into terminal effector cells and memory cells. IL-2 signals through a receptor complex composed of CD25/IL-2 R alpha, IL-2 R beta, and the Common gamma Chain (gamma c). IL-2 R beta is also a component of the IL-15 receptor complex. Gamma c is also a signaling subunit in the receptors for IL-4, -7, -9, -15, and -21.
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