Mouse/Rat TNF RI/TNFRSF1A Quantikine ELISA Kit

(25 citations)   
  • 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 (10 uL)
  • Sensitivity
    2.4 pg/mL
  • Assay Range
    15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum)
  • Specificity
    Natural and recombinant mouse and rat TNF RI
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity observed with 1 or more species tested.
  • Interference
    No significant interference observed with available related molecules.
Product Summary
The Quantikine Mouse/Rat TNF RI immunoassay is a 4.5 hour solid-phase ELISA designed to measure TNF RI in mouse or rat cell culture supernates and serum. It contains E. coli-expressed recombinant mouse TNF RI and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant mouse TNF RI. Results obtained using natural mouse or rat TNF RI showed dose response 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 TNF RI.

Precision
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.
Cell Culture Supernates, Serum
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean4611858148.3120592
Standard Deviation2.76.527.52.5633.3
CV%5.95.54.75.255.6

Recovery

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

Sample Type Average % Recovery Range %
Mouse Cell Culture Supernates (n=5) 97 89-109
Mouse Serum (n=5) 96 83-104
Rat Cell Culture Supernates (n=5) 98 92-116
Rat Serum (n=5) 96 88-104
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of TNF RI in each matrix were diluted with Calibrator Diluent and assayed.
Mouse/Rat TNF RI/TNFRSF1A Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: TNF RI/TNFRSF1A
TNF RI (TNF receptor 1; TNF R-p55/p60) is a transmembrane receptor for TNF-alpha and Lymphotoxin-alpha/TNF-beta. A soluble decoy form of the receptor is shed by ADAM-17/TACE in response to inflammatory stimulation. TNF RI signaling through NFkB is essential for the development of lymph node germinal centers and Peyer’s patches and for combating intracellular pathogens such as Listeria.
    • Long Name
      Tumor Necrosis Factor Receptor I
    • Entrez Gene IDs
      7132 (Human); 21937 (Mouse); 25625 (Rat); 102135285 (Cynomolgus Monkey);
    • Alternate Names
      CD120a antigen; CD120a; FPF; p55; p55-R; p60; TNF RI; TNFARMGC19588; TNF-R; TNF-R1; TNFR1TBP1; TNFR55; TNF-R55; TNFR60; TNFRI; TNF-RI; TNF-R-I; TNFR-I; TNFRSF1A; tumor necrosis factor binding protein 1; Tumor necrosis factor receptor 1; tumor necrosis factor receptor 1A isoform beta; tumor necrosis factor receptor superfamily member 1A; tumor necrosis factor receptor superfamily, member 1A; tumor necrosis factor receptor type 1; Tumor necrosis factor receptor type I; tumor necrosis factor-alpha receptor;
    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.
    Citations:

    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.

    25 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. Soluble TNF Regulates TACE via AP-2? Transcription Factor in Mouse Dendritic Cells
      Authors: L Ge, NL Vujanovic
      J. Immunol., 2017;198(1):417-427.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    2. Protective role of cellular prion protein against TNF?-mediated inflammation through TACE ?-secretase
      Authors: J Ezpeleta, F Boudet-Dev, M Pietri, A Baudry, V Baudouin, A Alleaume-B, N Dagoneau, O Kellermann, JM Launay, B Schneider
      Sci Rep, 2017;7(1):7671.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    3. Macrophage-derived tumor necrosis factor-alpha mediates diabetic renal injury.
      Authors: Awad A, You H, Gao T, Cooper T, Nedospasov S, Vacher J, Wilkinson P, Farrell F, Brian Reeves W
      Kidney Int, 2015;88(4):722-33.
    4. Kruppel-like factor 9 deficiency in uterine endometrial cells promotes ectopic lesion establishment associated with activated notch and hedgehog signaling in a mouse model of endometriosis.
      Authors: Heard M, Simmons C, Simmen F, Simmen R
      Endocrinology, 2014;155(4):1532-46.
      Species: Mouse
      Sample Type: Peritoneal Fluid
    5. Dose responsive effects of subcutaneous pentosan polysulfate injection in mucopolysaccharidosis type VI rats and comparison to oral treatment.
      Authors: Frohbergh M, Ge Y, Meng F, Karabul N, Solyom A, Lai A, Iatridis J, Schuchman E, Simonaro C
      PLoS ONE, 2014;9(6):e100882.
      Species: Rat
      Sample Type: Serum
    6. Expression of HIV transgene aggravates kidney injury in diabetic mice.
      Authors: Mallipattu S, Liu R, Zhong Y, Chen E, D'Agati V, Kaufman L, Ma'ayan A, Klotman P, Chuang P, He J
      Kidney Int, 2013;83(4):626-34.
      Species: Mouse
      Sample Type: Serum
    7. TNFR2 interposes the proliferative and NF-kappaB-mediated inflammatory response by podocytes to TNF-alpha.
      Authors: Bruggeman LA, Drawz PE, Kahoud N, Lin K, Barisoni L, Nelson PJ
      Lab. Invest., 2011;91(3):413-25.
      Species: Mouse
      Sample Type: Serum
    8. Restoration of regenerative osteoblastogenesis in aged mice: modulation of TNF.
      Authors: Wahl EC, Aronson J, Liu L, Fowlkes JL, Thrailkill KM, Bunn RC, Skinner RA, Miller MJ, Cockrell GE, Clark LM, Ou Y, Isales CM, Badger TM, Ronis MJ, Sims J, Lumpkin CK
      J. Bone Miner. Res., 2010;25(1):114-23.
      Species: Mouse
      Sample Type: Serum
    9. Ectodomain shedding of EGFR ligands and TNFR1 dictates hepatocyte apoptosis during fulminant hepatitis in mice.
      Authors: Murthy A, Defamie V, Smookler DS
      J. Clin. Invest., 2010;120(8):2731-44.
      Species: Mouse
      Sample Type: Serum
    10. Secretory lysosome targeting and induced secretion of human soluble TNF-alpha receptor in murine hematopoietic cells in vivo as a principle for immunoregulation in inflammation and malignancy.
      Authors: Johansson AC, Nandakumar KS, Persson AM, Olsson I, Hansson M
      Exp. Hematol., 2009;37(8):969-78.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    11. An eosinophil immune response characterizes the inflammatory skin disease observed in Tie-2 transgenic mice.
      Authors: Voskas D, Babichev Y, Ling LS, Alami J, Shaked Y, Kerbel RS, Ciruna B, Dumont DJ
      J. Leukoc. Biol., 2008;84(1):59-67.
      Species: Mouse
      Sample Type: Serum
    12. Hypoxia enhances lysosomal TNF-alpha degradation in mouse peritoneal macrophages.
      Authors: Lahat N, Rahat MA, Kinarty A, Weiss-Cerem L, Pinchevski S, Bitterman H
      Am. J. Physiol., Cell Physiol., 2008;295(1):C2-12.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    13. Fat diet and alcohol-induced steatohepatitis after LPS challenge in mice: role of bioactive TNF and Th1 type cytokines.
      Authors: Olleros ML, Martin ML, Vesin D, Fotio AL, Santiago-Raber ML, Rubbia-Brandt L, Spahr L, Hadengue A, Garcia I
      Cytokine, 2008;44(1):118-25.
      Species: Mouse
      Sample Type: Serum
    14. Differential roles of p55 and p75 tumor necrosis factor receptors on stretch-induced pulmonary edema in mice.
      Authors: Wilson MR, Goddard ME, O&amp;amp;apos;Dea KP, O'Dea KP, Choudhury S, Takata M
      Am. J. Physiol. Lung Cell Mol. Physiol., 2007;293(1):L60-8.
      Species: Mouse
      Sample Type: BALF
    15. Microparticles of human atherosclerotic plaques enhance the shedding of the tumor necrosis factor-alpha converting enzyme/ADAM17 substrates, tumor necrosis factor and tumor necrosis factor receptor-1.
      Authors: Canault M, Leroyer AS, Peiretti F, Leseche G, Tedgui A, Bonardo B, Alessi MC, Boulanger CM, Nalbone G
      Am. J. Pathol., 2007;171(5):1713-23.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    16. Bacterial translocation is downregulated by anti-TNF-alpha monoclonal antibody administration in rats with cirrhosis and ascites.
      Authors: Frances R, Chiva M, Sanchez E, Gonzalez-Navajas JM, Llovet T, Zapater P, Soriano G, Munoz C, Balanzo J, Perez-Mateo M, Song XY, Guarner C, Such J
      J. Hepatol., 2007;46(5):797-803.
      Species: Rat
      Sample Type: Serum
    17. The shedding activity of ADAM17 is sequestered in lipid rafts.
      Authors: Tellier E, Rebsomen L
      Exp. Cell Res., 2006;312(20):3969-80.
      Species: Mouse
      Sample Type: Tissue Homogenates
    18. FHL2 interacts with both ADAM-17 and the cytoskeleton and regulates ADAM-17 localization and activity.
      Authors: Canault M, Tellier E, Bonardo B, Mas E, Aumailley M, Juhan-Vague I, Nalbone G, Peiretti F
      J. Cell. Physiol., 2006;208(2):363-72.
      Species: Mouse
      Sample Type: Cell Lysates
    19. Role of tumour necrosis factor-alpha receptor p75 in cigarette smoke-induced pulmonary inflammation and emphysema.
      Authors: D'hulst AI, Bracke KR, Maes T, De Bleecker JL, Pauwels RA, Joos GF, Brusselle GG
      Eur. Respir. J., 2006;28(1):102-12.
      Species: Mouse
      Sample Type: BALF
    20. Nrf2 is a critical regulator of the innate immune response and survival during experimental sepsis.
      Authors: Thimmulappa RK, Lee H, Rangasamy T, Reddy SP, Yamamoto M, Kensler TW, Biswal S
      J. Clin. Invest., 2006;116(4):984-95.
      Species: Mouse
      Sample Type: Serum
    21. Systemic up-regulation of sTNFR2 and IL-6 in Porphyromonas gingivalis pneumonia in mice.
      Authors: Petelin M, Naruishi K, Shiomi N, Mineshiba J, Arai H, Nishimura F, Takashiba S, Murayama Y
      Exp. Mol. Pathol., 2004;76(1):76-81.
      Species: Human
      Sample Type: Serum
    22. A critical role for CCR2/MCP-1 interactions in the development of idiopathic pneumonia syndrome after allogeneic bone marrow transplantation.
      Authors: Hildebrandt GC, Duffner UA, Olkiewicz KM, Corrion LA, Willmarth NE, Williams DL, Clouthier SG, Hogaboam CM, Reddy PR, Moore BB, Kuziel WA, Liu C, Yanik G, Cooke KR
      Blood, 2004;103(6):2417-26.
      Species: Mouse
      Sample Type: BALF
    23. First-generation adenovirus vectors shorten survival time in a murine model of sepsis.
      Authors: Doerschug K, Sanlioglu S, Flaherty DM, Wilson RL, Yarovinsky T, Monick MM, Engelhardt JF, Hunninghake GW
      J. Immunol., 2002;169(11):6539-45.
      Species: Mouse
      Sample Type: Serum
    24. The protein tyrosine kinase inhibitor AG126 prevents the massive microglial cytokine induction by pneumococcal cell walls.
      Authors: Hanisch UK, Prinz M, Angstwurm K, Hausler KG, Kann O, Kettenmann H, Weber JR
      Eur. J. Immunol., 2001;31(7):2104-15.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    25. Enhanced murine macrophage TNF receptor shedding by cytosine-guanine sequences in oligodeoxynucleotides.
      Authors: Jin L, Raymond DP, Crabtree TD, Pelletier SJ, Houlgrave CW, Pruett TL, Sawyer RG
      J. Immunol., 2000;165(9):5153-60.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    Expand to show all 25 Citations
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