2 to 8 °C as supplied.
The dose response curve of the International Standard (87/586) parallels the OneStep standard curve. To convert sample values obtained with the kit to approximate NIBSC (87/586) units, use the following equation: NIBSC (87/586) approximate value (IU/mL) = 0.02 x OneStep Human IFNγ value (pg/mL).
Interferon gamma (IFN-γ) is a cytokine critical to both innate and adaptive immunity, and functions as the primary activator of macrophages, in addition to stimulating natural killer cells and neutrophils. A non-IgE-mediated anaphylactic reaction and severe bronchospasm have been reported once after the first injection of interferon gamma. IFN-γ activates cells via a different receptor than IFN-α and IFN-β, which accounts for the different physiological properties of the proteins. Production of IFN-γ is largely restricted to activated CD4+ TH1 T cells, CD8+ T cells, and natural killer cells. One of the most important consequences of IFN-γ secretion is the activation of macrophages. In addition, IFN-γ plays a central role in inflammatory responses by activating endothelial cells, promoting TH1 cell development and cellular immune responses, and up-regulation of major histocompatability complex protein expression on antigen-presenting cells.
Standard curve
Example of Human IFN-γ standard curve in Assay Diluent #10.
Standard curve
Example of NIBSC (87/586) standard curve in Assay Diluent #1. Human IFN-γ standard were calibrated by NIBSC (87/586) with a calibration factor (CF) of 1.0. Human IFN-γ standard concentration = NIBSC (87/586)/CF
Linearity
The concentrations of IFNγ were measured and interpolated from the target standard curves and corrected for sample dilution.
#1 sample is undiluted samples are as follows: human PBMC cells stimulated with 10ug/ml PHA for 5days (1%). The interpolated dilution factor corrected values are plotted. The mean target concentration was determined to be 31,555 pg/mL in stimulated human PBMC supernatant.
#2 sample is undiluted samples are as follows: human PBMC cells culture for 5days (50%). The interpolated dilution factor corrected values are plotted. The mean target concentration was determined to be 112 pg/mL in unstimulated human PBMC supernatant.
To assess the linearity of the assay, three samples were spiked with high concentrations of Human IFNγ in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
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