Porcine IFN-gamma Quantikine ELISA Kit

  (5 citations)     
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Product Details
Assay Procedure
Citations (5)
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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (100 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL)
  • Sensitivity
    11.2 pg/mL
  • Assay Range
    39.0 - 2,500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant porcine IFN-gamma
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Product Summary
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.

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
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 29 34 34
Mean 124 272 913 120 282 904
Standard Deviation 5.8 7.4 26.5 14 15.7 84.7
CV% 4.7 2.7 2.9 11.7 5.6 9.4

Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 36 32 36
Mean 226 449 1518 227 515 1522
Standard Deviation 10.1 12 40.4 17.1 51.3 97.7
CV% 4.5 2.7 2.7 7.5 10 6.4

Recovery

The recovery of porcine IFN-gamma spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=4) 97 84-116
EDTA Plasma (n=5) 98 85-111
Heparin Plasma (n=5) 106 86-118
Serum (n=5) 98 81-119
Linearity
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.
 IFN-gamma [Biotin]
Product Datasheets

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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.
  • Long Name:
    Interferon gamma
  • Entrez Gene IDs:
    3458 (Human); 15978 (Mouse); 25712 (Rat); 396991 (Porcine); 281237 (Bovine); 403801 (Canine); 493965 (Feline)
  • Alternate Names:
    IFG; IFI; IFNG; IFNgamma; IFN-gamma; Immune interferon; interferon gamma; interferon, gamma
Related Research Areas
Assay Procedure
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.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 100 µL Standard, Control, or Sample
  6.   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.
  7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

  8. 200 µL Conjugate
  9.   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.
  10.   Aspirate and wash 5 times.

  11. 200 µL Streptavidin-HRP
  12.   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.
  13.   Aspirate and wash 5 times.

  14. 120 µL Substrate Solution
  15. Add 120 µL Substrate Solution to each well.
  16. 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.

  17. 120 µL Stop Solution
  18. 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.
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

5 Citations: Showing 1 - 5
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Species
Sample Type
  1. Comparative measurement of cell-mediated immune responses of swine to the M and N proteins of porcine reproductive and respiratory syndrome virus.
    Authors: Jeong HJ, Song YJ, Lee SW
    Clin. Vaccine Immunol., 2010;17(4):503-12.
    Species: Porcine
    Sample Type: Cell Culture Supernates
  2. Metabolic activity of the enteric microbiota influences the fatty acid composition of murine and porcine liver and adipose tissues.
    Authors: Wall R, Ross RP, Shanahan F, O'Mahony L, O'Mahony C, Coakley M, Hart O, Lawlor P, Quigley EM, Kiely B, Fitzgerald GF, Stanton C
    Am. J. Clin. Nutr., 2009;89(5):1393-401.
    Species: Porcine
    Sample Type: Cell Culture Supernates
  3. Inhibition of complement and CD14 attenuates the Escherichia coli-induced inflammatory response in porcine whole blood.
    Authors: Thorgersen EB, Pharo A, Haverson K, Axelsen AK, Gaustad P, Kotwal GJ, Sfyroera G, Mollnes TE
    Infect. Immun., 2009;77(2):725-32.
    Species: Porcine
    Sample Type: Whole Blood
  4. Cytokine secretion depends on Galalpha(1,3)Gal expression in a pig-to-human whole blood model.
    Authors: Saethre M, Schneider MK, Lambris JD, Magotti P, Haraldsen G, Seebach JD, Mollnes TE
    J. Immunol., 2008;180(9):6346-53.
    Species: Porcine
    Sample Type: Cell Culture Supernates
  5. Blood concentrations of the cytokines IL-1beta, IL-6, IL-10, TNF-alpha and IFN-gamma during experimentally induced swine dysentery.
    Authors: Kruse R, Essen-Gustavsson B, Fossum C, Jensen-Waern M
    Acta Vet. Scand., 2008;50(0):32.
    Species: Porcine
    Sample Type: Serum

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