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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)
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Sensitivity
8 pg/mL
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Assay Range
15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma)
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Specificity
Natural and recombinant human 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.
Control Available
QC01-1, Quantikine Immunoassay Control Group 1 -
Product Summary
The Quantikine Human IFN-gamma Immunoassay is a 4.5 hour solid phase ELISA designed to measure IFN-gamma levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human IFN-gamma and antibodies raised against the recombinant factor. Results obtained for naturally occurring human IFN-gamma samples showed linear 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 human 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.
Cell Culture Supernates, Serum, EDTA Plasma | Intra-Assay Precision | Inter-Assay Precision |
Sample | 1 | 2 | 3 | 1 | 2 | 3 |
n | 20 | 20 | 20 | 40 | 40 | 40 |
Mean | 79.2 | 214 | 466 | 78.7 | 206 | 457 |
Standard Deviation | 3.7 | 5.5 | 13 | 6.1 | 13.1 | 17 |
CV% | 4.7 | 2.6 | 2.8 | 7.8 | 6.4 | 3.7 |
Recovery
The recovery of IFN-gamma spiked to levels throughout the range of the assay in various matrices was evaluated.
Sample Type | Average % Recovery | Range % |
Cell Culture Media (n=4) | 102 | 93-109 |
EDTA Plasma (n=5) | 98 | 88-111 |
Serum (n=5) | 102 | 91-118 |
Linearity
To assess the linearity of the assay, five samples were spiked with high concentrations of IFN-gamma in various matrices and diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
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