Mouse/Rat TNF RII/TNFRSF1B Quantikine ELISA Kit

(14 citations)
(1 Review)
  
  • 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), EDTA Plasma (10 uL)
  • Sensitivity
    5 pg/mL
  • Assay Range
    15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma)
  • Specificity
    Natural and recombinant mouse and rat TNF RII
  • 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 Mouse/Rat TNF RII Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse or rat TNF RII in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse TNF RII and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant protein accurately. Results obtained using natural mouse or rat TNF RII 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 mouse or rat TNF RII.

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, EDTA Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean37.211247238.6119500
Standard Deviation1.54.4211.43.514.7
CV%43.94.43.62.92.9

Recovery

The recovery of TNF RII 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) 102 83-119
Mouse EDTA Plasma (n=5) 95 80-119
Mouse Serum (n=5) 99 91-105
Rat Cell Culture Supernates (n=5) 98 85-116
Rat EDTA Plasma (n=5) 95 83-112
Rat Serum (n=5) 97 84-114
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of TNF RII were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Mouse/Rat TNF RII/TNFRSF1B Quantikine ELISA Kit
Mouse/Rat TNF RII/TNFRSF1B Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: TNF RII/TNFRSF1B
TNF RII (Tumor Necrosis Factor Receptor II), also known as TNFRSF1B, p75/p80, and CD120b, is a widely expressed receptor for membrane-associated TNF-alpha and Lymphotoxin-alpha. Its activation initiates pro-inflammatory and pro-survival responses via NFkB-dependent signaling pathways, although it may also induce apoptosis.
    • Long Name
      Tumor Necrosis Factor Receptor II
    • Entrez Gene IDs
      7133 (Human); 21938 (Mouse); 156767 (Rat);
    • Alternate Names
      CD120b antigen; CD120b; Etanercept; p75 TNF receptor; p75TBPII; p75TNFR; soluble TNFR1B variant 1; TNF RII; TNFBRp80 TNF-alpha receptor; TNF-R2; TNFR2TNFR1B; TNF-R75; TNFR80; TNFRII; TNF-RII; TNF-R-II; TNFR-II; TNFRSF1B; tumor necrosis factor beta receptor; tumor necrosis factor binding protein 2; Tumor necrosis factor receptor 2; tumor necrosis factor receptor superfamily member 1B; tumor necrosis factor receptor superfamily, member 1B; Tumor necrosis factor receptor type II;
    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.

    14 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. 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.
      Species: Mouse
      Sample Type: Tissue Homogenates
    2. 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
    3. 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
    4. 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
    5. 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
    6. 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
    7. 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
    8. 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
    9. 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
    10. 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
    11. 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
    12. 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: Mouse
      Sample Type: Serum
    13. 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
    14. 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 14 Citations
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