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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)
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Sensitivity
2 pg/mL
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Assay Range
9.4 - 600 pg/mL (Cell Culture Supernates, Serum)
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Specificity
Natural and recombinant mouse IFN-gamma
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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 Mouse IFN-gamma Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse IFN-gamma in cell culture supernates and mouse serum. It contains E. coli-expressed mouse IFN-gamma and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate recombinant mouse IFN-gamma accurately. Results obtained using natural mouse IFN-gamma 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 mouse IFN-gamma.
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.
Serum | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 45.1 | 143 | 528 | 42.8 | 139 | 518 |
| Standard Deviation | 2.1 | 4.4 | 11.8 | 3.6 | 13.6 | 46.5 |
| CV% | 4.7 | 3.1 | 2.2 | 8.4 | 9.8 | 9 |
Cell Culture Supernates | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 44.8 | 156 | 518 | 43.3 | 144 | 509 |
| Standard Deviation | 2.2 | 5 | 14.2 | 3.6 | 14.7 | 48.6 |
| CV% | 4.9 | 3.2 | 2.7 | 8.3 | 10.2 | 9.5 |
Recovery
The recovery of mouse IFN-gamma spiked to three levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Supernates (n=7) | 105 | 98-115 |
| Serum (n=5) | 97 | 91-106 |
Linearity
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of mouse IFN-gamma in each matrix were diluted with the appropriate Calibrator Diluent and then assayed.
Preparation and Storage
Background: IFN-gamma
IFN-gamma (Interferon-gamma) is the prototype proinflammatory cytokine and is produced by a variety of immune cells under inflammatory conditions, notably by T cells and NK cells. It plays a key role in host defense by promoting the development and activation of Th1 cells, chemoattraction and activation of monocytes and macrophages, upregulation of antigen presentation molecules, and immunoglobulin class switching in B cells. It also exhibits antiviral, antiproliferative, and apoptotic effects. In addition, IFN-gamma functions as an anti-inflammatory mediator by promoting the development of regulatory T cells and inhibiting Th17 cell differentiation. IFN-gamma dimers signal through a receptor complex of two IFN-gamma R1 and two IFN-gamma R2 subunits.
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Long Name
Interferon gamma
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