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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
3.5 hours
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Sample Type & Volume Required Per Well
Cell Culture Supernates (10 uL), Serum (13 uL), EDTA Plasma (13 uL), Heparin Plasma (13 uL)
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Sensitivity
10 pg/mL
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Assay Range
78.0 - 5,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Specificity
Natural and recombinant human IL-2 sR alpha
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Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
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Interference
No significant interference observed with available related molecules.
Control Available
QC05, Quantikine Immunoassay Control Group 2 -
Product Summary
The Quantikine Human IL-2 sR alpha Immunoassay is a 3.5 hour solid phase ELISA designed to measure IL-2 sR alpha in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human IL-2 sR alpha and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural human IL-2 sR alpha showed linear 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-2 sR alpha.
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 | 40 | 40 | 40 |
| Mean | 207 | 613 | 2357 | 188 | 630 | 2615 |
| Standard Deviation | 12.7 | 37.5 | 108 | 13.6 | 38.1 | 183 |
| CV% | 6.1 | 6.1 | 4.6 | 7.2 | 6 | 7 |
Recovery
The recovery of IL-2 sR alpha 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=5) | 105 | 93-118 |
| EDTA Plasma (n=5) | 103 | 91-113 |
| Heparin Plasma (n=5) | 104 | 98-113 |
| Serum (n=5) | 105 | 99-116 |
Linearity
To assess the linearity of the assay, five samples containing or spiked with high concentrations of IL-2 sR alpha in various matrices were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Background: CD25/IL-2 R alpha
CD25/IL-2 receptor alpha is a transmembrane protein that associates with IL-2 R beta/CD122 and the Common gamma Chain/CD132 on activated T cells and regulatory T cells (Treg). The Common gamma chain is a shared subunit in the receptors for IL-4, -7, -9, -15, and -21. IL-2 R alpha is required for mediating IL-2 induced effects including T cell proliferation, activation induced cell death (AICD) of autoreactive naive T cells, and the development of CD4+CD25+ Treg which contribute to peripheral T cell homeostasis. A soluble form of IL-2 R alpha can be generated by proteolytic cleavage of the cell surface receptor.
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