Canine IFN-gamma DuoSet ELISA

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Canine IFN-gamma ELISA Standard Curve
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Product Details
Citations (21)
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Canine IFN-gamma DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sample Volume Required
100 µL
Assay Range
31.2 - 2,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant canine IFN-gamma. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

Canine IFN-gamma ELISA Standard Curve

Product Datasheets

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Preparation and Storage

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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

Assay Procedure


Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Canine IFN-gamma DuoSet ELISA

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.

21 Citations: Showing 1 - 10
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  1. Immortalised canine buccal epithelial cells' CXCL8 secretion is affected by allergen extracts, Toll-like receptor ligands, IL-17A and calcitriol
    Authors: M Pelst, C Höbart, H de Rooster, B Devriendt, E Cox
    Veterinary research, 2022-09-13;53(1):72.
    Species: Canine
    Sample Types: Cell Culture Supernates
  2. Exploration of serum biomarkers in dogs with malignant melanoma receiving anti-PD-L1 therapy and potential of COX-2 inhibition for combination therapy
    Authors: N Maekawa, S Konnai, Y Asano, Y Sajiki, T Deguchi, T Okagawa, K Watari, H Takeuchi, S Takagi, K Hosoya, S Kim, H Ohta, Y Kato, Y Suzuki, S Murata, K Ohashi
    Oncogene, 2022-06-03;12(1):9265.
    Species: Canine
    Sample Types: Cell Culture Supernates
  3. miRNA-21 regulates CD69 and IL-10 expression in canine leishmaniasis
    Authors: JP Bragato, GT Rebech, JH Freitas, MOD Santos, SF Costa, FR Eugênio, PSPD Santos, VMF de Lima
    PLoS ONE, 2022-03-24;17(3):e0265192.
    Species: Canine
    Sample Types: cell culture supernatant
  4. Fluctuations in quality of life and immune responses during intravenous immunoglobulin infusion cycles
    Authors: JK Abbott, SK Chan, M MacBeth, JL Crooks, C Hancock, V Knight, EW Gelfand
    PLoS ONE, 2022-03-22;17(3):e0265852.
    Species: Canine
    Sample Types: cell culture supernatant
  5. Humoral Responses and Ex Vivo IFN-gamma Production after Canine Whole Blood Stimulation with Leishmania infantum Antigen or KMP11 Recombinant Protein
    Authors: P Martínez-O, N González, A Baldassarr, A Álvarez-Fe, L Ordeix, P Paradies, M Soto, L Solano-Gal
    Veterinary sciences, 2022-03-04;9(3):.
    Species: Canine
    Sample Types: Serum
  6. Comparison of circulating CD4+, CD8+ lymphocytes and cytokine profiles between dogs with atopic dermatitis and healthy dogs
    Authors: MT Verde, S Villanueva, A Loste, D Marteles, D Pereboom, T Conde, A Fernández
    Research in Veterinary Science, 2022-01-29;145(0):13-20.
    Species: Canine
    Sample Types: Serum
  7. A pilot clinical study of the therapeutic antibody against canine PD-1 for advanced spontaneous cancers in dogs
    Authors: M Igase, Y Nemoto, K Itamoto, K Tani, M Nakaichi, M Sakurai, Y Sakai, S Noguchi, M Kato, T Tsukui, T Mizuno
    Sci Rep, 2020-10-27;10(1):18311.
    Species: Canine
    Sample Types: Cell Culture Supernates
  8. Evaluation of canine leishmaniosis vaccine CaniLeish� under field conditions in native dog populations from an endemic Mediterranean area - a randomized controlled trial
    Authors: R Velez, E Domenech, A Rodríguez-, D Barrios, S Tebar, A Fernández-, R Aguilar, C Dobaño, J Alberola, J Cairó, M Gállego
    Acta Trop., 2020-02-05;0(0):105387.
    Species: Canine
    Sample Types: Cell Culture Supernates
  9. Canine non-B, non-T NK lymphocytes have a potential antibody-dependent cellular cytotoxicity function against antibody-coated tumor cells
    Authors: Y Kim, SH Lee, CJ Kim, JJ Lee, D Yu, S Ahn, DJ Shin, SK Kim
    BMC Vet. Res., 2019-10-14;15(1):339.
    Species: Canine
    Sample Types: Cell Culture Supernates
  10. Local immune and microbiological responses to mucosal administration of a Liposome-TLR agonist immunotherapeutic in dogs
    Authors: W Wheat, L Chow, A Kuzmik, S Soontarara, J Kurihara, M Lappin, S Dow
    BMC Vet. Res., 2019-09-13;15(1):330.
    Species: Canine
    Sample Types: Cell Culture Supernates
  11. IL12 p35 and p40 subunit genes administered as pPAL plasmid constructs do not improve protection of pPAL-LACK vaccine against canine leishmaniasis
    Authors: PJ Alcolea, A Alonso, A Esteban, P Peris, A Cortés, JA Castillo, V Larraga
    PLoS ONE, 2019-02-22;14(2):e0212136.
    Species: Canine
    Sample Types: Serum
  12. PD-1 and PD-L1 regulate cellular immunity in canine visceral leishmaniasis
    Authors: KL Oliveira S, V Marin Chik, G Luvizotto, AA Correa Lea, BF de Almeida, F De Rezende, PSP Dos Santos, G Fabrino Ma, VMF De Lima
    Comp. Immunol. Microbiol. Infect. Dis., 2018-12-14;62(0):76-87.
    Species: Canine
    Sample Types: Cell Culture Supernates
  13. Mechanisms of Immune Suppression Utilized by Canine Adipose and Bone Marrow-Derived Mesenchymal Stem Cells
    Stem Cells Dev., 2017-01-24;0(0):.
    Species: Canine
    Sample Types: Cell Culture Supernates
  14. Impact of LbSapSal Vaccine in Canine Immunological and Parasitological Features before and after Leishmania chagasi-Challenge
    PLoS ONE, 2016-08-24;11(8):e0161169.
    Species: Canine
    Sample Types: Cell Culture Supernates
  15. Evaluation of Live Recombinant Nonpathogenic Leishmania tarentolae Expressing Cysteine Proteinase and A2 Genes as a Candidate Vaccine against Experimental Canine Visceral Leishmaniasis.
    Authors: Shahbazi M, Zahedifard F, Taheri T, Taslimi Y, Jamshidi S, Shirian S, Mahdavi N, Hassankhani M, Daneshbod Y, Zarkesh-Esfahani S, Papadopoulou B, Rafati S
    PLoS ONE, 2015-07-21;10(7):e0132794.
    Species: Canine
    Sample Types: Cell Culture Supernates
  16. One-year timeline kinetics of cytokine-mediated cellular immunity in dogs vaccinated against visceral leishmaniasis.
    Authors: Costa-Pereira C, Moreira M, Soares R, Marteleto B, Ribeiro V, Franca-Dias M, Cardoso L, Viana K, Giunchetti R, Martins-Filho O, Araujo M
    BMC Vet Res, 2015-04-11;11(0):92.
    Species: Canine
    Sample Types: Cell Culture Supernates
  17. Expression of regulatory T cells in jejunum, colon, and cervical and mesenteric lymph nodes of dogs naturally infected with Leishmania infantum.
    Authors: Figueiredo M, Deoti B, Amorim I, Pinto A, Moraes A, Carvalho C, da Silva S, de Assis A, de Faria A, Tafuri W
    Infect Immun, 2014-06-16;82(9):3704-12.
    Species: Canine
    Sample Types: Tissue Homogenates
  18. Expression of PD-L1 on canine tumor cells and enhancement of IFN-gamma production from tumor-infiltrating cells by PD-L1 blockade.
    Authors: Maekawa N, Konnai S, Ikebuchi R, Okagawa T, Adachi M, Takagi S, Kagawa Y, Nakajima C, Suzuki Y, Murata S, Ohashi K
    PLoS ONE, 2014-06-10;9(6):e98415.
    Species: Canine
    Sample Types: Cell Culture Supernates
  19. Testing the efficacy of a multi-component DNA-prime/DNA-boost vaccine against Trypanosoma cruzi infection in dogs.
    Authors: Aparicio-Burgos JE, Ochoa-Garcia L, Zepeda-Escobar JA, Gupta S, Dhiman M, Martinez JS, de Oca-Jimenez RM, Val Arreola M, Barbabosa-Pliego A, Vazquez-Chagoyan JC, Garg NJ
    PLoS Negl Trop Dis, 2011-05-17;5(5):e1050.
    Species: Canine
    Sample Types: Serum
  20. Clinical, parasitologic, and immunologic evolution in dogs experimentally infected with sand fly-derived Leishmania chagasi promastigotes.
    Authors: Travi BL, Osorio EY, Saldarriaga OA
    Am. J. Trop. Med. Hyg., 2009-12-01;81(6):994-1003.
    Species: Canine
    Sample Types: Cell Culture Supernates
  21. Protective immunity against challenge with Leishmania (Leishmania) chagasi in beagle dogs vaccinated with recombinant A2 protein.
    Authors: Fernandes AP, Costa MM, Coelho EA, Michalick MS, de Freitas E, Melo MN, Luiz Tafuri W, Resende Dde M, Hermont V, Abrantes Cde F, Gazzinelli RT
    Vaccine, 2008-09-09;26(46):5888-95.
    Species: Canine
    Sample Types: Cell Culture Supernates


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Canine IFN-gamma DuoSet ELISA
By Kumudhini Haran on 06/26/2020
Sample Tested: T cells