Canine IFN-gamma Quantikine ELISA Kit

Catalog # Availability Size / Price Qty
CAIF00
Canine IFN-gamma ELISA Linearity
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Product Details
Procedure
Citations (11)
FAQs
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Reviews (5)

Canine IFN-gamma Quantikine ELISA Kit Summary

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
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 111 459 1234 119 457 1315
Standard Deviation 10.1 35.1 92.4 10.1 21.5 64.8
CV% 9.1 7.6 7.5 8.5 4.7 4.9

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 ELISA Linearity

Data Examples

Canine IFN-gamma ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

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 for Canine IFN-gamma Quantikine ELISA Kit

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.

11 Citations: Showing 1 - 10
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  1. Development of canine PD-1/PD-L1 specific monoclonal antibodies and amplification of canine T cell function
    Authors: JW Choi, SS Withers, H Chang, JA Spanier, VL De La Trin, H Panesar, BT Fife, R Sciammas, EE Sparger, PF Moore, MS Kent, RB Rebhun, SJ McSorley
    PLoS ONE, 2020;15(7):e0235518.
    Species: Dog
    Sample Types: Cell Culture Supernates
  2. Oat beta-glucan as a dietary supplement for dogs
    Authors: LG Ferreira, M Endrighi, KG Lisenko, MRD de Oliveir, MR Damasceno, JA Claudino, PG Gutierres, AP Peconick, FMOB Saad, MG Zangeronim
    PLoS ONE, 2018;13(7):e0201133.
    Species: Canine
    Sample Types: Plasma
  3. Lymphocytic, cytokine and transcriptomic profiles in peripheral blood of dogs with atopic dermatitis
    BMC Vet Res, 2016;12(1):174.
    Species: Canine
    Sample Types: Plasma
  4. 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 Types: Serum
  5. 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 Types: Serum
  6. 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 Types: Cell Culture Supernates
  7. 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 Types: Cell Culture Supernates
  8. 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 Types: Cell Culture Supernates
  9. 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 Types: Serum
  10. 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 Types: Cell Culture Supernates
  11. 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 Types: Cell Culture Supernates

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Reviews for Canine IFN-gamma Quantikine ELISA Kit

Average Rating: 5 (Based on 5 Reviews)

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Canine IFN-gamma Quantikine ELISA Kit
By Anonymous on 06/16/2020
Sample Tested: cell supernatant from cultured T cells

Canine IFN-gamma Quantikine ELISA Kit
By Anonymous on 05/26/2020
Sample Tested: Cell Culture Media

The standards in this kit work very well - over 0.99 R squared every time I use it. Great assay. Would recommend to a colleague. Consistent and reliable.


Canine IFN-gamma Quantikine ELISA Kit
By Anonymous on 03/10/2020
Sample Tested: Peripheral blood mononuclear cells (PBMCs)

Canine IFN-gamma Quantikine ELISA Kit
By Leslie Priddy on 04/03/2018
Sample Tested: Serum

Canine IFN-gamma Quantikine ELISA Kit
By chen xie on 12/01/2017
Sample Tested: Serum and Plasma