Porcine TNF-alpha Quantikine ELISA Kit

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  • 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
    5 pg/mL
  • Assay Range
    23.4 - 1,500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant porcine 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 Porcine TNF-alpha Immunoassay is a 4.5 hour solid phase ELISA designed to measure porcine TNF-alpha in cell culture supernates, serum, and plasma samples. It contains E. coli-expressed recombinant porcine TNF-alpha and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately measure recombinant porcine TNF-alpha. Results obtained using natural porcine TNF-alpha show dose response curves that are 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 porcine TNF-alpha.

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested 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.
Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Standard Deviation7.213369.73272

Cell Culture Supernates
Intra-Assay Precision Inter-Assay Precision
Standard Deviation4.99.2278.82444


The recovery of porcine TNF-alpha 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) 99 91-105
EDTA Plasma (n=6) 98 83-111
Heparin Plasma (n=4) 91 82-99
Serum (n=6) 105 91-120
To assess the linearity of the assay, six samples containing and/or spiked with various concentrations of porcine TNF-alpha in each matrix were diluted with the appropriate Calibrator Diluent and then assayed.
Porcine TNF-alpha Quantikine ELISA Kit
Preparation and Storage
  • 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); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit);
    • Alternate Names
      APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; tnfa; tnf-a; TNFalpha; TNF-alphacachectin; TNFATNF, macrophage-derived; 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;
    Related Research Areas
    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. 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 2 hours.
    10.   Aspirate and wash 5 times.

    11. 100 µL Substrate Solution
    12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

    13. 100 µL Stop Solution
    14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

    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.

    Showing Results 1 - 10 of 31
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    Sample Type
    1. Inhalation therapy with the synthetic TIP-like peptide AP318 attenuates pulmonary inflammation in a porcine sepsis model.
      Authors: Hartmann E, Ziebart A, Thomas R, Liu T, Schad A, Tews M, Moosmann B, Kamuf J, Duenges B, Thal S, David M
      BMC Pulm Med, 2015;15(0):7.
      Species: Porcine
      Sample Type: Plasma
    2. Efficacy of linezolid compared to vancomycin in an experimental model of pneumonia induced by methicillin-resistant Staphylococcus aureus in ventilated pigs.
      Authors: Martinez-Olondris P, Rigol M, Soy D, Guerrero L, Agusti C, Quera MA, Li Bassi G, Esperatti M, Luque N, Liapikou M, Filella X, Marco F, de la Bellacasa JP, Torres A
      Crit. Care Med., 2012;40(1):162-8.
      Species: Porcine
      Sample Type: Serum
    3. Risks Associated with High-Dose Lactobacillus rhamnosus in an Escherichia coli Model of Piglet Diarrhoea: Intestinal Microbiota and Immune Imbalances.
      Authors: Li XQ, Zhu YH, Zhang HF, Yue Y, Cai ZX, Lu QP, Zhang L, Weng XG, Zhang FJ, Zhou D, Yang JC, Wang JF
      PLoS ONE, 2012;7(7):e40666.
      Species: Porcine
      Sample Type: Serum
    4. Ornithodoros moubata complement inhibitor is an equally effective C5 inhibitor in pigs and humans.
      Authors: Barratt-Due A, Thorgersen EB, Lindstad JK, Pharo A, Lissina O, Lambris JD, Nunn MA, Mollnes TE
      J. Immunol., 2011;187(9):4913-9.
      Species: Porcine
      Sample Type: Plasma
    5. Plasma concentrations of inflammatory cytokines rise rapidly during ECMO-related SIRS due to the release of preformed stores in the intestine.
      Authors: McILwain RB, Timpa JG, Kurundkar AR
      Lab. Invest., 2010;90(1):128-39.
      Species: Porcine
      Sample Type: Plasma
    6. CD14 inhibition efficiently attenuates early inflammatory and hemostatic responses in Escherichia coli sepsis in pigs.
      Authors: Thorgersen EB, Hellerud BC, Nielsen EW, Barratt-Due A, Fure H, Lindstad JK, Pharo A, Fosse E, Tonnessen TI, Johansen HT, Castellheim A, Mollnes TE
      FASEB J., 2010;24(3):712-22.
      Species: Porcine
      Sample Type: Plasma
    7. Dissecting the effects of lipopolysaccharides from nonlipopolysaccharide molecules in experimental porcine meningococcal sepsis.
      Authors: Hellerud BC, Nielsen EW, Thorgersen EB, Lindstad JK, Pharo A, Tonnessen TI, Castellheim A, Mollnes TE, Brandtzaeg P
      Crit. Care Med., 2010;38(6):1467-74.
      Species: Porcine
      Sample Type: Plasma
    8. Metabolic activity of the enteric microbiota influences the fatty acid composition of murine and porcine liver and adipose tissues.
      Authors: Wall R, Ross RP, Shanahan F, O'Mahony L, O'Mahony C, Coakley M, Hart O, Lawlor P, Quigley EM, Kiely B, Fitzgerald GF, Stanton C
      Am. J. Clin. Nutr., 2009;89(5):1393-401.
      Species: Porcine
      Sample Type: Cell Culture Supernates
    9. Cardiac Function and the Proinflammatory Cytokine Response After Recovery From Cardiac Arrest in Swine.
      Authors: Niemann JT, Rosborough JP, Youngquist S, Shah AP, Lewis RJ, Phan QT, Filler SG
      J. Interferon Cytokine Res., 2009;0(0):.
      Species: Porcine
      Sample Type: Plasma
    10. Autologous mesenchymal stem cells produce reverse remodelling in chronic ischaemic cardiomyopathy.
      Authors: Schuleri KH, Feigenbaum GS, Centola M, Weiss ES, Zimmet JM, Turney J, Kellner J, Zviman MM, Hatzistergos KE, Detrick B, Conte JV, McNiece I, Steenbergen C, Lardo AC, Hare JM
      Eur. Heart J., 2009;30(22):2722-32.
      Species: Porcine
      Sample Type: Serum
    11. Effect of linezolid compared with glycopeptides in methicillin-resistant Staphylococcus aureus severe pneumonia in piglets.
      Authors: Luna CM, Bruno DA, Garcia-Morato J, Mann KC, Risso Patron J, Sagardia J, Absi R, Garcia Bottino M, Marchetti D, Famiglietti A, Baleztena M, Biancolini C
      Chest, 2009;135(6):1564-71.
      Species: Porcine
      Sample Type: Serum
    12. Inhibition of complement and CD14 attenuates the Escherichia coli-induced inflammatory response in porcine whole blood.
      Authors: Thorgersen EB, Pharo A, Haverson K, Axelsen AK, Gaustad P, Kotwal GJ, Sfyroera G, Mollnes TE
      Infect. Immun., 2009;77(2):725-32.
      Species: Porcine
      Sample Type: Whole Blood
    13. Blood concentrations of the cytokines IL-1beta, IL-6, IL-10, TNF-alpha and IFN-gamma during experimentally induced swine dysentery.
      Authors: Kruse R, Essen-Gustavsson B, Fossum C, Jensen-Waern M
      Acta Vet. Scand., 2008;50(0):32.
      Species: Porcine
      Sample Type: Serum
    14. Effects of glucocorticoids in ventilated piglets with severe pneumonia.
      Authors: Sibila O, Luna CM, Agusti C, Baquero S, Gando S, Patron JR, Morato JG, Absi R, Bassi N, Torres A
      Eur. Respir. J., 2008;32(4):1037-46.
      Species: Porcine
      Sample Type: Serum
    15. Protective effect of Sivelestat in a porcine hepatectomy model prepared using an intermittent Pringle method.
      Authors: Shimoda M, Iwasaki Y, Okada T, Sawada T, Kubota K
      Eur. J. Pharmacol., 2008;587(1):248-52.
      Species: Porcine
      Sample Type: Serum
    16. Inducible nitric oxide synthase expression and cardiomyocyte dysfunction during sustained moderate ischemia in pigs.
      Authors: Heinzel FR, Gres P, Boengler K, Duschin A, Konietzka I, Rassaf T, Snedovskaya J, Meyer S, Skyschally A, Kelm M, Heusch G, Schulz R
      Circ. Res., 2008;103(10):1120-7.
      Species: Porcine
      Sample Type: Cell Lysates
    17. Cytokine secretion depends on Galalpha(1,3)Gal expression in a pig-to-human whole blood model.
      Authors: Saethre M, Schneider MK, Lambris JD, Magotti P, Haraldsen G, Seebach JD, Mollnes TE
      J. Immunol., 2008;180(9):6346-53.
      Species: Porcine
      Sample Type: Cell Culture Supernates
    18. Experimental Pseudomonas aeruginosa pneumonia: evaluation of the associated inflammatory response.
      Authors: Sibila O, Agusti C, Torres A, Baquero S, Gando S, Patron JR, Morato JG, Goffredo DH, Bassi N, Luna CM
      Eur. Respir. J., 2007;30(6):1167-72.
      Species: Porcine
      Sample Type: Serum
    19. Involvement of both tumor necrosis factor-alpha-induced necrosis and p53-mediated caspase-dependent apoptosis in nephrotoxicity of cisplatin.
      Authors: Yano T, Itoh Y, Matsuo M, Kawashiri T, Egashira N, Oishi R
      Apoptosis, 2007;12(10):1901-9.
      Species: Porcine
      Sample Type: Cell Culture Supernates
    20. Activated protein C downregulates p38 mitogen-activated protein kinase and improves clinical parameters in an in-vivo model of septic shock.
      Authors: Nold MF, Nold-Petry CA, Fischer D, Richter B, Blaheta R, Pfeilschifter J, Muhl H, Schranz D, Veldman A
      Thromb. Haemost., 2007;98(5):1118-26.
      Species: Porcine
      Sample Type: Plasma
    21. Bidirectional role of tumor necrosis factor-alpha in coronary microembolization: progressive contractile dysfunction versus delayed protection against infarction.
      Authors: Skyschally A, Gres P, Hoffmann S, Haude M, Erbel R, Schulz R, Heusch G
      Circ. Res., 2007;100(1):140-6.
      Species: Porcine
      Sample Type: Tissue Homogenates
    22. Experimental severe Pseudomonas aeruginosa pneumonia and antibiotic therapy in piglets receiving mechanical ventilation.
      Authors: Luna CM, Baquero S, Gando S, Patron JR, Morato JG, Sibila O, Absi R, Famiglietti A, Vay CA, Von Stecher F, Agusti C, Torres A
      Chest, 2007;132(2):523-31.
      Species: Porcine
      Sample Type: Serum
    23. The fibrin-derived peptide Bbeta15-42 is cardioprotective in a pig model of myocardial ischemia-reperfusion injury.
      Authors: Roesner JP, Petzelbauer P, Koch A, Mersmann J, Zacharowski PA, Boehm O, Reingruber S, Pasteiner W, Mascher D, Wolzt M, Barthuber C, Noldge-Schomburg GE, Scheeren TW, Zacharowski K
      Crit. Care Med., 2007;35(7):1730-5.
      Species: Porcine
      Sample Type: Plasma
    24. Proangiogenic effects of protease-activated receptor 2 are tumor necrosis factor-alpha and consecutively Tie2 dependent.
      Authors: Zhu T, Sennlaub F, Beauchamp MH, Fan L, Joyal JS, Checchin D, Nim S, Lachapelle P, Sirinyan M, Hou X, Bossolasco M, Rivard GE, Heveker N, Chemtob S
      Arterioscler. Thromb. Vasc. Biol., 2006;26(4):744-50.
      Species: Porcine
      Sample Type: Cell Culture Supernates
    25. Role of oxidative stress in remodeling of the myocardial microcirculation in hypertension.
      Authors: Zhu XY, Daghini E, Chade AR, Rodriguez-Porcel M, Napoli C, Lerman A, Lerman LO
      Arterioscler. Thromb. Vasc. Biol., 2006;26(8):1746-52.
      Species: Porcine
      Sample Type: Plasma
    26. Dietary fatty acid composition rather than vitamin E supplementation influence ex vivo cytokine and eicosanoid response of porcine alveolar macrophages.
      Authors: Moller S, Lauridsen C
      Cytokine, 2006;35(1):6-12.
      Species: Porcine
      Sample Type: Cell Culture Supernates
    27. Conjugated linoleic acid attenuates the production and gene expression of proinflammatory cytokines in weaned pigs challenged with lipopolysaccharide.
      Authors: Changhua L, Jindong Y, Defa L, Lidan Z, Shiyan Q, Jianjun X
      J. Nutr., 2005;135(2):239-44.
      Species: Porcine
      Sample Type: Serum
    28. The effect of warm humidified CO2 on the dissipation of residual gas following laparoscopy in piglets.
      Authors: Glew PA, Campher MJ, Pearson K, Schofield JC, Davey AK
      J Am Assoc Gynecol Laparosc, 2004;11(2):204-10.
      Species: Porcine
      Sample Type: Serum
    29. Deciphering the involvement of innate immune factors in the development of the host response to PRRSV vaccination.
      Authors: Royaee AR, Husmann RJ, Dawson HD, Calzada-Nova G, Schnitzlein WM, Zuckermann FA, Lunney JK
      Vet. Immunol. Immunopathol., 2004;102(3):199-216.
      Species: Porcine
      Sample Type: Whole Cells
    30. Extracellular ubiquitin inhibits the TNF-alpha response to endotoxin in peripheral blood mononuclear cells and regulates endotoxin hyporesponsiveness in critical illness.
      Authors: Majetschak M, Krehmeier U, Bardenheuer M, Denz C, Quintel M, Voggenreiter G, Obertacke U
      Blood, 2003;101(5):1882-90.
      Species: Porcine
      Sample Type: Plasma
    31. Ex vivo adenoviral vector gene delivery results in decreased vector-associated inflammation pre- and post-lung transplantation in the pig.
      Authors: Yeung JC, Wagnetz D, Cypel M, Rubacha M, Koike T, Chun YM, Hu J, Waddell TK, Hwang DM, Liu M, Keshavjee S
      Mol. Ther., 212;20(6):1204-11.
      Species: Porcine
      Sample Type: Plasma
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