Mouse/Rat TNF RII/TNFRSF1B Quantikine ELISA Kit

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MRT20
Mouse/Rat TNF RII/TNFRSF1B ELISA Standard Curve
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Citations (14)
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Mouse/Rat TNF RII/TNFRSF1B Quantikine ELISA Kit Summary

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
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 37.2 112 472 38.6 119 500
Standard Deviation 1.5 4.4 21 1.4 3.5 14.7
CV% 4 3.9 4.4 3.6 2.9 2.9

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 ELISA Linearity
Mouse/Rat TNF RII/TNFRSF1B ELISA Linearity

Data Examples

Mouse/Rat TNF RII/TNFRSF1B ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
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
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

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 for Mouse/Rat TNF RII/TNFRSF1B Quantikine 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.

14 Citations: Showing 1 - 10
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  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 Types: 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 Types: 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 Types: Serum
  4. 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 Types: Cell Culture Supernates
  5. 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 Types: Serum
  6. 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 Types: BALF
  7. 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 Types: Serum
  8. 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 Types: Tissue Homogenates
  9. 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 Types: BALF
  10. 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 Types: Serum
  11. 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 Types: Cell Lysates
  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 Types: 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 Types: 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 Types: Cell Culture Supernates

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Mouse/Rat TNF RII/TNFRSF1B Quantikine ELISA Kit
By Anonymous on 05/02/2017
Sample Tested: Cell Culture Media