Canine IFN-gamma Quantikine ELISA Kit

(9 citations)
(1 Review)
  
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
  • Sensitivity
    60 pg/mL
  • Assay Range
    62.5 - 4,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant canine 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 Canine IFN-gamma Immunoassay is a 4 hour solid-phase ELISA designed to measure canine IFN-gamma in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant canine IFN-gamma and antibodies raised against the recombinant factor. Results obtained for naturally occurring canine 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 canine IFN-gamma.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times 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
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean11145912341194571315
Standard Deviation10.135.192.410.121.564.8
CV%9.17.67.58.54.74.9

Recovery

The recovery of canine 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) 103 92-111
EDTA Plasma (n=4) 96 86-102
Heparin Plasma (n=4) 98 93-106
Serum (n=4) 96 83-107
Linearity
To assess the linearity of the assay, samples containing or spiked with high concentrations of canine IFN-gamma in each matrix were diluted with Calibrator Diluent and assayed.
Canine IFN-gamma Quantikine ELISA Kit
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. 50 µL Standard, Control, or sample
    6.   Add 50 µL of Standard, Control, or sample to each well. Tap the plate gently for 1 minute. Cover with a plate sealer, and incubate at room temperature for 2 hours.
    7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

    8. 100 µL Conjugate
    9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour.
    10.   Aspirate and wash 5 times.

    11. 100 µL Streptavidin-HRP
    12.   Add 100 µL Streptavidin-HRP to each well. Cover with a new plate sealer, and incubate at room temperature for 30 minutes.
    13.   Aspirate and wash 5 times.

    14. 100 µL Substrate Solution
    15. Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

    16. 100 µL Stop Solution
    17. Add 100 µL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing. 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.

    9 Citations: Showing 1 - 9
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    Species
    Sample Type
    1. Lymphocytic, cytokine and transcriptomic profiles in peripheral blood of dogs with atopic dermatitis
      BMC Vet Res, 2016;12(1):174.
      Species: Canine
      Sample Type: Plasma
    2. Defining the Pharmacodynamic Profile and Therapeutic Index of NHS-IL12 Immunocytokine in Dogs with Malignant Melanoma.
      Authors: Paoloni M, Mazcko C, Selting K, Lana S, Barber L, Phillips J, Skorupski K, Vail D, Wilson H, Biller B, Avery A, Kiupel M, LeBlanc A, Bernhardt A, Brunkhorst B, Tighe R, Khanna C
      PLoS ONE, 2015;10(6):e0129954.
      Species: Canine
      Sample Type: Serum
    3. Use of a recombinant cysteine proteinase from Leishmania (Leishmania) infantum chagasi for the Immunotherapy of canine visceral Leishmaniasis.
      Authors: Ferreira J, Silva L, Longo-Maugeri I, Katz S, Barbieri C
      PLoS Negl Trop Dis, 2014;8(3):e2729.
      Species: Canine
      Sample Type: Serum
    4. Identification of myeloid derived suppressor cells in dogs with naturally occurring cancer.
      Authors: Goulart MR, Pluhar GE, Ohlfest JR
      PLoS ONE, 2012;7(3):e33274.
      Species: Canine
      Sample Type: Cell Culture Supernates
    5. Human Flt3L generates dendritic cells from canine peripheral blood precursors: implications for a dog glioma clinical trial.
      Authors: Xiong W, Candolfi M, Liu C
      PLoS ONE, 2010;5(6):e11074.
      Species: Canine
      Sample Type: Cell Culture Supernates
    6. Prime-boost immunization with DNA followed by a recombinant vaccinia virus expressing P50 induced protective immunity against Babesia gibsoni infection in dogs.
      Authors: Fukumoto S, Tamaki Y, Okamura M, Bannai H, Yokoyama N, Suzuki T, Igarashi I, Suzuki H, Xuan X
      Vaccine, 2007;25(7):1334-41.
      Species: Canine
      Sample Type: Serum
    7. Injection of a recombinant AAV serotype 2 into canine skeletal muscles evokes strong immune responses against transgene products.
      Authors: Yuasa K, Yoshimura M, Urasawa N, Ohshima S, Howell JM, Nakamura A, Hijikata T, Miyagoe-Suzuki Y, Takeda S
      Gene Ther., 2007;14(17):1249-60.
      Species: Canine
      Sample Type: Cell Culture Supernates
    8. A functional comparison of canine and murine bone marrow derived cultured mast cells.
      Authors: Lin TY, London CA,
      Vet. Immunol. Immunopathol., 2006;114(3):320-34.
      Species: Canine
      Sample Type: Cell Culture Supernates
    9. Effect of ovarian hormones on periodical changes in immune resistance associated with estrous cycle in the beagle bitch.
      Authors: Sugiura K, Nishikawa M, Ishiguro K, Tajima T, Inaba M, Torii R, Hatoya S, Wijewardana V, Kumagai D, Tamada H, Sawada T, Ikehara S, Inaba T
      Immunobiology, 2004;209(8):619-27.
      Species: Canine
      Sample Type: Cell Culture Supernates
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