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Canine TNF-alpha Quantikine ELISA Kit

Catalog #: CATA00
18 Citations 1 Review Datasheet / COA / SDS
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CATA00
Canine TNF-alpha ELISA Standard Curve
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Citations (18)
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Reviews (1)
Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Canine TNF-alpha Quantikine ELISA 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 (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
Sensitivity
4.2 pg/mL
Assay Range
7.8 - 500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant canine TNF-alpha
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 TNF-alpha Immunoassay is a 4.5 hour solid-phase ELISA designed to measure canine TNF-alpha in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant canine TNF-alpha and antibodies raised against the recombinant protein. Results obtained for naturally occurring canine TNF-alpha 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 TNF-alpha.

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 20 100 66 106
Mean (pg/mL) 18.6 123 201 20.9 54 149
Standard Deviation 1.5 6.5 15.4 2.1 5.2 13.8
CV% 8.1 5.3 7.7 10 9.6 9.3

Recovery

The recovery of canine TNF-alpha spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=7) 99 83-109
EDTA Plasma (n=6) 94 82-103
Heparin Plasma (n=6) 94 88-103
Serum (n=6) 92 83-106

Linearity

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

Scientific Data

N/A Canine TNF-alpha ELISA Standard Curve View Larger

Canine TNF-alpha ELISA Standard Curve

Product Datasheets

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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: TNF-alpha

Tumor necrosis factor alpha (TNF-α), also known as cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, apoptosis, and immune system development. TNF-α is produced by a wide variety of immune and epithelial cell types. Human TNF-α consists of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD). Within the ECD, human TNF-α shares 97% aa sequence identity with rhesus and 71% - 92% with bovine, canine, cotton rat, equine, feline, mouse, porcine, and rat TNF-α. The 26 kDa type 2 transmembrane protein is assembled intracellularly to form a noncovalently linked homotrimer. Ligation of this complex induces reverse signaling that promotes lymphocyte costimulation but diminishes monocyte responsiveness.

Cleavage of membrane bound TNF-α by TACE/ADAM17 releases a 55 kDa soluble trimeric form of TNF-α. TNF-α trimers bind the ubiquitous TNF RI and the hematopoietic cell-restricted TNF RII, both of which are also expressed as homotrimers. TNF-α regulates lymphoid tissue development through control of apoptosis. It also promotes inflammatory responses by inducing the activation of vascular endothelial cells and macrophages. TNF-α is a key cytokine in the development of several inflammatory disorders. It contributes to the development of type 2 diabetes through its effects on insulin resistance and fatty acid metabolism.

Long Name:
Tumor Necrosis Factor alpha
Entrez Gene IDs:
7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 102139631 (Cynomolgus Monkey); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)
Alternate Names:
APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; TNFA; TNF-A; TNFalpha; TNF-alpha; TNF-alphacachectin; TNFATNF, macrophage-derived; TNFG1F; TNFSF1A; TNFSF2; TNFSF2TNF superfamily, member 2; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor; tumor necrosis factor-alpha
&#9888; 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
  3.   Add 50 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or sample
  4.   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.
  5.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
    6. 100 µL Conjugate
  6.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour.
  7.   Aspirate and wash 5 times.
    8. 100 µL Streptavidin-HRP
  8.   Add 100 µL Streptavidin-HRP to each well. Cover with a new plate sealer, and incubate at room temperature for 30 minutes.
  9.   Aspirate and wash 5 times.
    10. 100 µL Substrate Solution
  10. Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    11. 100 µL Stop Solution
  11. 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 TNF-alpha 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.

18 Citations: Showing 1 - 10
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  1. Phase I/II Trial of Urokinase Plasminogen Activator-Targeted Oncolytic Newcastle Disease Virus for Canine Intracranial Tumors
    Authors: Rossmeisl, JH;King, JN;Robertson, JL;Weger-Lucarelli, J;Elankumaran, S;
    Cancers
    Species: Canine
    Sample Types: Serum
  2. Effects of SKCPT on Osteoarthritis in Beagle Meniscectomy and Cranial Cruciate Ligament Transection Models
    Authors: Kim, HM;Kang, M;Jung, YS;Lee, YJ;Choi, W;Yoo, H;Kim, J;An, HJ;
    International journal of molecular sciences
    Species: Canine
    Sample Types: Synovial Fluid
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  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, 2020-10-05;0(0):.
    Species: Canine
    Sample Types: Plasma
  9. A preliminary study assessing cytokine production following ex vivo stimulation of whole blood with diet in dogs with chronic enteropathy
    Authors: A Kathrani, E Hall
    BMC Vet. Res., 2019-06-04;15(1):185.
    Species: Canine
    Sample Types: Cell Culture Supernates
  10. Clinical Assessment of Intravenous Endothelial Progenitor Cell Transplantation in Dogs
    Authors: SH Lee, JC Ra, HJ Oh, MJ Kim, EMN Setyawan, YB Choi, JW Yang, SK Kang, SH Han, GA Kim, BC Lee
    Cell Transplant, 2019-04-24;0(0):9636897188216.
    Species: Canine
    Sample Types: Serum
  11. Evaluation of the in vitro effect of Boldo and Meadowsweet plant extracts on the expression of antimicrobial peptides and inflammatory markers in canine keratinocytes
    Authors: D Santoro, K Ahrens, R Vesny, C Navarro, H Gatto, R Marsella
    Res. Vet. Sci., 2017-05-17;115(0):255-262.
    Species: Canine
    Sample Types: Cell Culture Supernates
  12. 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., 2017-04-13;13(1):104.
    Species: Canine
    Sample Types: Plasma
  13. The effect of intra-articular botulinum toxin A on substance P, prostaglandin E2, and tumor necrosis factor alpha in the canine osteoarthritic joint
    Authors: HM Heikkilä, AK Hielm-Björ, JF Innes, OM Laitinen-V
    BMC Vet. Res., 2017-03-21;13(1):74.
    Species: Canine
    Sample Types: Serum
  14. Outcomes of Spatially Fractionated Radiotherapy (GRID) for Bulky Soft Tissue Sarcomas in a Large Animal Model
    Authors: MW Nolan, TL Gieger, AA Karakashia, MN Nikolova-K, LP Posner, DM Roback, JN Rivera, S Chang
    Technol. Cancer Res. Treat, 2017-02-07;0(0):1533034617690.
    Species: Canine
    Sample Types: Serum
  15. 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
  16. TLR5 risk-associated haplotype for canine inflammatory bowel disease confers hyper-responsiveness to flagellin.
    Authors: Kathrani A, Holder A, Catchpole B, Alvarez L, Simpson K, Werling D, Allenspach K
    PLoS ONE, 2012-01-18;7(1):e30117.
    Species: Canine
    Sample Types: Serum
  17. 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
  18. A functional comparison of canine and murine bone marrow derived cultured mast cells.
    Authors: Lin TY, London CA
    Vet. Immunol. Immunopathol., 2006-10-06;114(3):320-34.
    Species: Canine
    Sample Types: Cell Culture Supernates

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