• Assay Length
    5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (100 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL), Citrate Plasma (100 uL)
  • Sensitivity
    0.736 pg/mL
  • Assay Range
    2.2 - 7,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
  • Specificity
    Natural and recombinant human TNF-alpha
  • Cross-reactivity
    Cross-reactivity observed with 1 or more available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    Interference observed with 1 or more available related molecules.
Control Available
QC196, QuantiGlo Immunoassay Control Set 732 for Human TNF-alpha - Please Inquire
Product Summary
The QuantiGlo Human TNF-alpha Chemiluminscent Immunoassay is a 5 hour solid phase ELISA designed to measure TNF-alpha in cell culture supernates, serum, and plasma. It contains E. coli-derived recombinant human TNF-alpha and antibodies raised against the recombinant factor. It has been shown to accurately quantitate recombinant human TNF-alpha. Results obtained using naturally occurring TNF-alpha samples showed linear curves that were parallel to the standard curves obtained using the QuantiGlo kit standards. These results indicate that this kit can be used to determine relative mass values for natural TNF-alpha.

Intra-Assay Precision (Precision within an assay) Four samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Four samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates
Intra-Assay Precision Inter-Assay Precision
Standard Deviation0.341.3612.61320.472.928.1313

Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma
Intra-Assay Precision Inter-Assay Precision
Standard Deviation0.441.5114.41530.423.1224.6244


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

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 107 97-115
Citrate Plasma (n=4) 100 91-109
EDTA Plasma (n=4) 100 86-112
Heparin Plasma (n=4) 100 83-114
Serum (n=4) 101 87-113
To assess the linearity of the assay, samples spiked with high concentrations of TNF-alpha in various matrices were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human TNF-alpha QuantiGlo 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-alpha; 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

    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.

    15 Citations: Showing 1 - 10
    Filter your results:

    Sample Type
    1. Orai and TRPC channel characterization in Fc?RI-mediated calcium signaling and mediator secretion in human mast cells
      Authors: HE Wajdner, J Farrington, C Barnard, PT Peachell, CG Schnackenb, JP Marino, X Xu, K Affleck, M Begg, EP Seward
      Physiol Rep, 2017;5(5):.
      Species: Human
      Sample Type: Cell Culture Supernates
    2. Early B cell factor 1 regulates adipocyte morphology and lipolysis in white adipose tissue.
      Authors: Gao H, Mejhert N, Fretz J, Arner E, Lorente-Cebrian S, Ehrlund A, Dahlman-Wright K, Gong X, Stromblad S, Douagi I, Laurencikiene J, Dahlman I, Daub C, Ryden M, Horowitz M, Arner P
      Cell Metab, 2014;19(6):981-92.
      Species: Human
      Sample Type: Cell Culture Supernates
    3. Effect of renal impairment on cognitive function during a 3-year follow up in elderly patients with type 2 diabetes: Association with microinflammation.
      Authors: Kawamura T, Umemura T, Umegaki H, Imamine R, Kawano N, Tanaka C, Kawai M, Minatoguchi M, Kusama M, Kouchi Y, Watarai A, Kanai A, Nakashima E, Hotta N
      J Diabetes Investig, 2014;5(5):597-605.
      Species: Human
      Sample Type: Plasma
    4. Calcium sensing receptor activation elevates proinflammatory factor expression in human adipose cells and adipose tissue.
      Authors: Cifuentes M, Fuentes C, Tobar N, Acevedo I, Villalobos E, Hugo E, Ben-Jonathan N, Reyes M
      Mol Cell Endocrinol, 2012;361(1):24-30.
      Species: Human
      Sample Type: Cell Culture Supernates
    5. Dexamethasone and lenalidomide have distinct functional effects on erythropoiesis.
      Authors: Narla A, Dutt S, McAuley JR
      Blood, 2011;118(8):2296-304.
      Species: Human
      Sample Type: Cell Culture Supernates
    6. Effect of intensive lifestyle changes on endothelial function and on inflammatory markers of atherosclerosis.
      Authors: Dod HS, Bhardwaj R, Sajja V, Weidner G, Hobbs GR, Konat GW, Manivannan S, Gharib W, Warden BE, Nanda NC, Beto RJ, Ornish D, Jain AC
      Am. J. Cardiol., 2010;105(3):362-7.
      Species: Human
      Sample Type: Serum
    7. Dexamethasone suppresses interleukin-22 associated with bacterial infection in vitro and in vivo.
      Authors: Ziesche E, Scheiermann P, Bachmann M, Sadik CD, Hofstetter C, Zwissler B, Pfeilschifter J, Muhl H
      Clin. Exp. Immunol., 2009;157(3):370-6.
      Species: Human
      Sample Type: Plasma
    8. Obesity in pregnancy stimulates macrophage accumulation and inflammation in the placenta.
      Authors: Challier JC, Basu S, Bintein T, Minium J, Hotmire K, Catalano PM, Hauguel-De Mouzon S
      Placenta, 2008;29(3):274-81.
      Species: Human
      Sample Type: Plasma
    9. Autocrine tumor necrosis factor alpha links endoplasmic reticulum stress to the membrane death receptor pathway through IRE1alpha-mediated NF-kappaB activation and down-regulation of TRAF2 expression.
      Authors: Hu P, Han Z, Couvillon AD, Kaufman RJ, Exton JH
      Mol. Cell. Biol., 2006;26(8):3071-84.
      Species: Human
      Sample Type: Cell Culture Supernates
    10. Effect of influenza vaccine on markers of inflammation and lipid profile.
      Authors: Tsai MY, Hanson NQ, Straka RJ, Hoke TR, Ordovas JM, Peacock JM, Arends VL, Arnett DK
      J. Lab. Clin. Med., 2005;145(6):323-7.
      Species: Human
      Sample Type: Plasma
    11. Possible contribution of adipocytokines on diabetic neuropathy.
      Authors: Matsuda M, Kawasaki F, Inoue H, Kanda Y, Yamada K, Harada Y, Saito M, Eto M, Matsuki M, Kaku K
      Diabetes Res. Clin. Pract., 2004;66(0):S121-3.
      Species: Human
      Sample Type: Plasma
    12. Impact of fibronectin fragments on the transendothelial migration of HIV-infected leukocytes and the development of subendothelial foci of infectious leukocytes.
      Authors: Birdsall HH, Porter WJ, Green DM, Rubio J, Trial J, Rossen RD
      J. Immunol., 2004;173(4):2746-54.
      Species: Human
      Sample Type: Cell Culture Supernates
    13. A potential pro-angiogenic cell therapy with human placenta-derived mesenchymal cells.
      Authors: Nishishita T, Ouchi K, Zhang X, Inoue M, Inazawa T, Yoshiura K, Kuwabara K, Nakaoka T, Watanabe N, Igura K, Takahashi TA, Yamashita N
      Biochem. Biophys. Res. Commun., 2004;325(1):24-31.
      Species: Human
      Sample Type: Cell Lysates
    14. Plasma MMP-2 and MMP-9 and their inhibitors TIMP-1 and TIMP-2 during human orthotopic liver transplantation. The effect of aprotinin and the relation to ischemia/reperfusion injury.
      Authors: Kuyvenhoven JP, Molenaar IQ, Verspaget HW, Veldman MG, Palareti G, Legnani C, Moolenburgh SE, Terpstra OT, Lamers CB, van Hoek B, Porte RJ
      Thromb. Haemost., 2004;91(3):506-13.
      Species: Human
      Sample Type: Plasma
    15. Procalcitonin and proinflammatory cytokine clearance during continuous venovenous haemofiltration in septic patients.
      Authors: Dahaba AA, Elawady GA, Rehak PH, List WF
      Anaesth Intensive Care, 2002;30(3):269-74.
      Species: Human
      Sample Type: Plasma
    Expand to show all 15 Citations
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