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

Name: Canine IL-6 Quantikine Kit
Brand: R&D Systems
SKU: CA6000
Rating: 4.5 (2 reviews)

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.

Canine IL-6 Quantikine Kit

Catalog #: CA6000
17 Citations 2 Reviews Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
CA6000

Best Seller

2 Images

All IL-6 ELISAs

Product Details

Procedure

Citations (17)

FAQs

Supplemental Products

Reviews (2)

Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Canine IL-6 Quantikine Kit Summary

Assay Type

Solid Phase Sandwich ELISA

Format

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)

Sensitivity

11.8 pg/mL

Assay Range

31.3 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)

Specificity

Natural and recombinant canine IL-6

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 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.

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	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

Recovery

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

Linearity

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.

Scientific Data

N/A

View Larger

Canine IL-6 ELISA Standard Curve

Product Datasheets

Product Datasheet

COA

SDS

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: 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

Related Research Areas

⚠ 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.

Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

50 µL Assay Diluent

Add 50 µL of Assay Diluent to each well.

100 µL Standard, Control, or sample

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.
Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

200 µL Conjugate

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.
Aspirate and wash 5 times.

120 µL Substrate Solution

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

120 µL Stop Solution

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.

17 Citations: Showing 1 - 10
Filter your results:

Filter by:

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, 2023-04-06;18(4):e0273138.
Species: Canine
Sample Types: Serum
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
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, 2022-10-02;9(10):.
Species: Canine
Sample Types: Cell Culture Supernates
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, 2022-07-12;12(1):11818.
Species: Canine
Sample Types: Cell Culture Supernates
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, 2021-12-08;0(0):.
Species: Canine
Sample Types: Serum
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, 2021-11-05;0(0):.
Species: Canine
Sample Types: Urine
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, 2021-09-04;8(9):.
Species: Canine
Sample Types: Whole Blood
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, 2021-04-26;17(1):176.
Species: Canine
Sample Types: Serum
Effect of modified ultrafiltration on cytokines and hemoconcentration in dogs undergoing cardiopulmonary bypass
Authors: H Suzuki, N Oshima, T Watari
J Vet Med Sci, 2020-10-05;0(0):.
Species: Canine
Sample Types: Plasma
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, 2018-03-08;5(0):343-347.
Species: Canine
Sample Types: Serum

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., 2018-02-27;0(0):.
Species: Canine
Sample Types: Serum

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., 2017-07-17;0(0):.
Species: Canine
Sample Types: Serum

Higher neonatal growth rate and body condition score at 7ï¿½months 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., 2017-04-13;13(1):104.
Species: Canine
Sample Types: Plasma

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, 2016-03-31;24(1):40.
Species: Canine
Sample Types: Plasma

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, 2014-09-30;16(1):75-85.
Species: Canine
Sample Types: CSF

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, 2014-09-06;10(0):194.
Species: Canine
Sample Types: Serum

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, 2010-06-11;5(6):e11074.
Species: Canine
Sample Types: Cell Culture Supernates

FAQs

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.

View all ELISA FAQs