Human TNF-alpha QuantiGlo ELISA Kit

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QTA00B
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Human TNF-alpha Chemiluminescent ELISA Calibrator Diluent RD5P (1X) Standard Curve
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Citations (16)
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Human TNF-alpha QuantiGlo ELISA Kit Summary

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.

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.

Precision

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

Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 4 1 2 3 4
n 20 20 20 20 20 20 20 20
Mean 5.17 37.7 413 3678 5.97 37.4 382 3869
Standard Deviation 0.44 1.51 14.4 153 0.42 3.12 24.6 244
CV% 8.5 4 3.5 4.2 7 8.3 6.4 6.3

Cell Culture Supernates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 4 1 2 3 4
n 20 20 20 20 20 20 20 20
Mean 5.94 39.7 390 3636 6.34 38 374 3711
Standard Deviation 0.34 1.36 12.6 132 0.47 2.9 28.1 313
CV% 5.7 3.4 3.2 3.6 7.4 7.6 7.5 8.4

Recovery

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

Linearity

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 Chemiluminescent ELISA Linearity

Data Examples

Human TNF-alpha Chemiluminescent ELISA Calibrator Diluent RD5P (1X) Standard Curve

Human TNF-alpha Chemiluminescent ELISA Calibrator Diluent RD6N Standard Curve

Product Datasheets

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

Citations for Human TNF-alpha QuantiGlo 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.

16 Citations: Showing 1 - 10
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  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 Types: Cell Culture Supernates
  2. Elevated Serum Level of HMGB1 in Patients with the Antiphospholipid Syndrome
    Authors: V Manganelli, A Capozzi, S Truglia, C Alessandri, E Lococo, T Garofalo, C De Carolis, F Conti, G Valesini, M Sorice, A Longo, R Misasi
    J Immunol Res, 2017;2017(0):4570715.
    Species: Human
    Sample Types: Serum
  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 Types: Plasma
  4. 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 Types: Cell Culture Supernates
  5. 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 Types: Cell Culture Supernates
  6. 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 Types: Cell Culture Supernates
  7. 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 Types: Serum
  8. 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 Types: Plasma
  9. 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 Types: Plasma
  10. 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 Types: Cell Culture Supernates
  11. 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 Types: Plasma
  12. 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 Types: Cell Lysates
  13. 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 Types: Plasma
  14. 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 Types: Plasma
  15. 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 Types: Cell Culture Supernates
  16. 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 Types: Plasma

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