< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
No significant interference observed with available related molecules.
The Quantikine Porcine IFN-gamma Immunoassay is a 4 hour solid-phase ELISA designed to measure porcine IFN-gamma in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant porcine IFN-gamma and antibodies raised against the recombinant factor. Results obtained for naturally occurring porcine IFN-gamma 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 porcine IFN-gamma.
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
The recovery of porcine IFN-gamma spiked to levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Supernates (n=4)
EDTA Plasma (n=5)
Heparin Plasma (n=5)
To assess the linearity of the assay, samples containing or spiked with high concentrations of porcine IFN-gamma in each matrix were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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.
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
50 µL Assay Diluent
Add 50 µL of Assay Diluent to each well.
100 µL Standard, Control, or Sample
Add 100 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
200 µL Conjugate
Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour on the shaker.
Aspirate and wash 5 times.
200 µL Streptavidin-HRP
Add 200 µL Streptavidin-HRP to each well. Cover with a new plate sealer, and incubate at room temperature for 30 minutes on the shaker.
Aspirate and wash 5 times.
120 µL Substrate Solution
Add 120 µL Substrate Solution to each well.
For Serum & Plasma Samples: Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT. For Cell Culture Supernate Samples: Incubate at room temperature for 20 minutes on the benchtop. PROTECT FROM LIGHT.
120 µL Stop Solution
Add 120 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
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but also provides information about sample types, species, and experimental conditions.