Canine IL-6 Quantikine Kit

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Canine IL-6 ELISA Standard Curve
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Product Details
Citations (16)
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Canine IL-6 Quantikine Kit Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (100 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL)
11.8 pg/mL
Assay Range
31.3 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Natural and recombinant canine IL-6
< 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.

Product Summary

The Quantikine Canine IL-6 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure canine IL-6 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant canine IL-6 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural canine IL-6 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 naturally occurring canine IL-6.


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 67 65 66
Mean (pg/mL) 92.6 256 685 90.7 262 666
Standard Deviation 3.1 8.3 15.3 6.6 15.5 30.2
CV% 3.3 3.2 2.2 7.3 5.9 4.5


The recovery of canine IL-6 spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=6) 94 87-103
EDTA Plasma (n=4) 97 91-103
Heparin Plasma (n=4) 92 83-100
Serum (n=4) 93 84-102


To assess the linearity of the assay, samples containing or spiked with various concentrations of canine IL-6 in each matrix were diluted with Calibrator Diluent and then assayed.
Canine IL-6 ELISA Linearity

Scientific Data

Canine IL-6 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.
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: IL-6

Interleukin 6 (IL-6) is a pleiotropic, alpha -helical, 22-28 kDa phosphorylated and variably glycosylated cytokine that plays important roles in the acute phase reaction, inflammation, hematopoiesis, bone metabolism, and cancer progression (1-5). Mature canine IL-6 is 187 amino acids (aa) in length and shares 58%, 37%, 39%, and 76% aa sequence identity with human, mouse, rat, and feline IL-6, respectively. Cells known to express IL-6 include CD8+ T cells, fibroblasts, synoviocytes, adipocytes, osteoblasts, megakaryocytes, endothelial cells (under the influence of endothelins), sympathetic neurons, cerebral cortex neurons, adrenal medulla chromaffin cells, retinal pigment cells, mast cells, keratinocytes, Langerhans cells, fetal and adult astrocytes, neutrophils, monocytes, eosinophils, colonic epithelial cells, B1 B cells and pancreatic islet beta cells (2, 6-28). IL-6 production is generally correlated with cell activation and is normally kept in control by glucocorticoids, catecholamines, and secondary sex steroids (2). Normal human circulating IL-6 is in the 1 pg/mL range, with slight elevations during the menstrual cycle, modest elevations in certain cancers, and large elevations after surgery (29-33). 

IL-6 induces signaling through a cell surface heterodimeric receptor complex composed of a ligand binding subunit (IL-6 R alpha) and a signal transducing subunit (gp130). IL-6 binds to IL-6 R alpha, triggering IL-6 R alpha association with gp130 and gp130 dimerization (34). gp130 is also a component of the receptors for CLC, CNTF, CT-1, IL-11, IL-27, LIF, and OSM (35). Soluble forms of IL-6 R alpha are generated by both alternative splicing and proteolytic cleavage (5). In a mechanism known as trans-signaling, complexes of soluble IL-6 and IL-6 R alpha elicit responses from gp130- expressing cells that lack cell surface IL-6 R alpha (5). Trans-signaling enables a wider range of cell types to respond to IL-6, as the expression of gp130 is ubiquitous, while that of IL-6 R alpha is predominantly restricted to hepatocytes, monocytes, and resting lymphocytes (2, 5). Soluble splice forms of gp130 block trans-signaling from IL-6/IL-6 R alpha but not from other cytokines that use gp130 as a co-receptor (5, 36). 
IL-6, along with TNF-alpha and IL-1, drives the acute inflammatory response. IL-6 is almost solely responsible for fever and the acute phase response in the liver, and it is important in the transition from acute inflammation to either acquired immunity or chronic inflammatory disease (1-5). When dysregulated, it contributes to chronic inflammation in conditions such as obesity, insulin resistance, inflammatory bowel disease, arthritis, and sepsis (2, 5). IL-6 modulates bone resorption and is a major effector of inflammatory joint destruction in rheumatoid arthritis through its promotion of Th17 cell development and activity (1). It contributes to atherosclerotic plaque development and destabilization as well as the development of inflammation-associated carcinogenesis (1, 2). IL-6 can also function as an anti-inflammatory molecule, as in skeletal muscle where it is secreted in response to exercise (2). In addition, it enhances hematopoietic stem cell proliferation and the differentiation of memory B cells and plasma cells (37). 

Long Name:
Interleukin 6
Entrez Gene IDs:
3569 (Human); 16193 (Mouse); 24498 (Rat); 399500 (Porcine); 280826 (Bovine); 403985 (Canine); 102138971 (Cynomolgus Monkey); 100034196 (Equine); 493687 (Feline); 463288 (Primate); 100008733 (Rabbit)
Alternate Names:
B-cell differentiation factor; B-cell stimulatory factor 2; BSF2; BSF-2; CDF; CTL differentiation factor ; HSF; hybridoma growth factor; IFNB2; IFN-beta-2; IL6; IL-6; Interferon beta-2; interleukin 6 (interferon, beta 2); interleukin BSF-2; interleukin-6; MGI-2A
⚠ 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

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 an 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 2 hours on the shaker.
  10.   Aspirate and wash 5 times.

  11. 120 µL Substrate Solution
  12.   Add 120 µL Substrate Solution to each well. Incubate at room temperature on the benchtop for 30 minutes. PROTECT FROM LIGHT.

  13. 120 µL Stop Solution
  14.   Add 120 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 IL-6 Quantikine 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.

16 Citations: Showing 1 - 10
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  1. Translation of immunomodulatory therapy to treat chronic heart failure: Preclinical studies to first in human
    Authors: HD Humes, KD Aaronson, DA Buffington, HN Sabbah, AJ Westover, LT Yessayan, B Szamosfalv, FD Pagani
    PLoS ONE, 2023;18(4):e0273138.
    Species: Canine
    Sample Types: Serum
  2. Molecular Characterization of CF33 Canine Cell Line and Evaluation of Its Ability to Respond against Infective Stressors in Sight of Anticancer Approaches
    Authors: E Razzuoli, CG De Ciucis, B Chirullo, K Varello, R Zoccola, L Guardone, P Petrucci, D Rubini, E Bozzetta, M Goria, F Fruscione, P Modesto
    Veterinary sciences, 2022;9(10):.
    Species: Canine
    Sample Types: Cell Culture Supernates
  3. Mycobacterium intracellulare induces a Th17 immune response via M1-like macrophage polarization in canine peripheral blood mononuclear cells
    Authors: S Kim, YS Hyun, HT Park, MK Shin, HS Yoo
    Scientific Reports, 2022;12(1):11818.
    Species: Canine
    Sample Types: Cell Culture Supernates
  4. Effect of Cannabidiol (CBD) on Canine Inflammatory Response: An Ex Vivo Study on LPS Stimulated Whole Blood
    Authors: E Gugliandol, P Licata, AF Peritore, R Siracusa, R D'Amico, M Cordaro, R Fusco, D Impellizze, R Di Paola, S Cuzzocrea, R Crupi, CD Interlandi
    Veterinary sciences, 2021;8(9):.
    Species: Canine
    Sample Types: Whole Blood
  5. Inflammation and its association with oxidative stress in dogs with heart failure
    Authors: A Nemec Svet, B Verk, N ?ebulj-Kad, J Salobir, V Rezar, A Domanjko P
    Bmc Veterinary Research, 2021;17(1):176.
    Species: Canine
    Sample Types: Serum
  6. Prospective evaluation of 5 urinary biomarkers as predictors of acute kidney injury in nonazotemic, hospitalized dogs
    Authors: R Nivy, N Chaim, E Hanael, GA Sutton, Y Bruchim, I Aroch, G Segev
    Journal of veterinary internal medicine, 2021;0(0):.
    Species: Canine
    Sample Types: Urine
  7. Interleukin-6 and thrombopoietin concentrations in dogs with carcinoma with and without thrombocytosis
    Authors: A Cheney, AD Woolcock, A Mukhopadhy, D Knapp, GE Moore
    Journal of veterinary internal medicine, 2021;0(0):.
    Species: Canine
    Sample Types: Serum
  8. Effect of modified ultrafiltration on cytokines and hemoconcentration in dogs undergoing cardiopulmonary bypass
    Authors: H Suzuki, N Oshima, T Watari
    J Vet Med Sci, 2020;0(0):.
    Species: Canine
    Sample Types: Plasma
  9. Hematologic and serologic status of military working dogs given standard diet containing natural botanical supplements
    Authors: E Lee, JH Choi, HJ Jeong, SG Hwang, S Lee, JW Oh
    Toxicol Rep, 2018;5(0):343-347.
    Species: Canine
    Sample Types: Serum
  10. Evaluation of potential serum biomarkers of hepatic fibrosis and necroinflammatory activity in dogs with liver disease
    Authors: C Raghu, J Ekena, JM Cullen, CB Webb, LA Trepanier
    J. Vet. Intern. Med., 2018;0(0):.
    Species: Canine
    Sample Types: Serum
  11. Higher neonatal growth rate and body condition score at 7�months are predictive factors of obesity in adult female Beagle dogs
    Authors: L Leclerc, C Thorin, J Flanagan, V Biourge, S Serisier, P Nguyen
    BMC Vet. Res., 2017;13(1):104.
    Species: Canine
    Sample Types: Plasma
  12. Comparison of postoperative pain and inflammation reaction in dogs undergoing preventive laparoscopic-assisted and incisional gastropexy
    Authors: T Haraguchi, S Kimura, H Itoh, S Nishikawa, M Hiyama, K Tani, T Iseri, Y Itoh, M Nakaichi, Y Taura, K Itamoto
    J. Vet. Med. Sci., 2017;0(0):.
    Species: Canine
    Sample Types: Serum
  13. Continuous venovenous hemofiltration decreases mortality and ameliorates acute lung injury in canine model of severe salt water drowning
    Authors: J Chen, G Chen, D Xiao, W Peng, G Yu, Y Lin, F Zheng
    Scand J Trauma Resusc Emerg Med, 2016;24(1):40.
    Species: Canine
    Sample Types: Plasma
  14. Canine model of ischemic stroke with permanent middle cerebral artery occlusion: clinical features, magnetic resonance imaging, histopathology, and immunohistochemistry.
    Authors: Jeon J, Jung H, Jang H, Moon J, Park K, Lee H, Lim H, Sur J, Kang B, Ha J, Jung D
    J Vet Sci, 2015;16(1):75-85.
    Species: Canine
    Sample Types: CSF
  15. Evaluation of the effects of methadone and tramadol on postoperative analgesia and serum interleukin-6 in dogs undergoing orthopaedic surgery.
    Authors: Cardozo L, Cotes L, Kahvegian M, Rizzo M, Otsuki D, Ferrigno C, Fantoni D
    BMC Vet Res, 2014;10(0):194.
    Species: Canine
    Sample Types: Serum
  16. 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


  1. What is the normal appearance of Assay Diluent RD1-75?

    • Assay Diluent RD1-75 contains high concentrations of additives and typically has a very cloudy, brown appearance. It is recommended to mix this diluent well before and during use.

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Canine IL-6 Quantikine ELISA Kit
By chen xie on 12/01/2017
Sample Tested: Serum and Plasma

Canine IL-6 Quantikine ELISA Kit
By Adam Guess on 10/14/2016
Sample Tested: Cell Culture Supernates