Mouse IL-1ra/IL-1F3 Quantikine ELISA Kit MRA00: R&D Systems

Mouse IL-1ra/IL-1F3 Quantikine ELISA Kit

(22 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 (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
  • Sensitivity
    13 pg/mL
  • Assay Range
    31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant mouse IL-1ra
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    Interference observed with 1 or more available related molecules.
Product Summary
The Quantikine Mouse IL-1ra Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse IL-1ra in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse IL-1ra and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse IL-1ra 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 naturally occurring mouse IL-1ra.

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, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020262627
Mean632271226662251233
Standard Deviation5.112.247.26.112.529.9
CV%8.15.43.89.25.62.4

Recovery

The recovery of mouse IL-1ra spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=5) 110 95-120
EDTA Plasma (n=4) 92 82-101
Heparin Plasma (n=4) 91 83-101
Serum (n=4) 95 88-104
Linearity
To assess the linearity of the assay, samples containing or spiked with high concentrations of mouse IL-1ra in each matrix were diluted with Calibrator Diluent and then assayed.
Mouse IL-1ra/IL-1F3 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: IL-1ra/IL-1F3
IL-1ra (IL-1 receptor antagonist) is a widely expressed acute phase protein that serves to dampen inflammation. IL-1ra binds to both IL-1 RI and IL-1 RII. It competitively inhibits IL-1 (alpha or beta) binding to IL-1 RI and does not trigger recruitment of the accessory molecule IL-1 R AcP or activation of IL-1 RI signaling. IL-1ra blocks the proinflammatory actions of IL-1 including Prostaglandin E2 and IL-6 release from endothelial cells, fever generation, thrombocytosis, the production of hepatic acute phase proteins, and neutrophil expansion and infiltration.
    • Long Name
      Interleukin 1 Receptor Antagonist
    • Entrez Gene IDs
      3557 (Human); 16181 (Mouse); 60582 (Rat); 397499 (Porcine);
    • Alternate Names
      DIRA; ICIL-1RA; IL1F3; IL-1F3; IL1ra; IL-1ra; IL-1ra3; IL1RN; IL-1RN; interleukin 1 receptor antagonist; IRAP; MVCD4;
    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 on a horizontal orbital microplate shaker.
    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 on the shaker.
    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 on the benchtop. 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.

    22 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. IL-24 Promotes Pseudomonas aeruginosa Keratitis in C57BL/6 Mouse Corneas
      Authors: BX Ross, N Gao, X Cui, TJ Standiford, J Xu, FX Yu
      J. Immunol, 2017;0(0):.
      Species: Mouse
      Sample Type: Tissue Homogenates
    2. Controlled Release of Interleukin-1 Receptor Antagonist from Hyaluronic Acid-Chitosan Microspheres Attenuates Interleukin-1?-Induced Inflammation and Apoptosis in Chondrocytes
      Biomed Res Int, 2016;2016(0):6290957.
    3. C-type Lectin Mincle Recognizes Glucosyl-diacylglycerol of Streptococcus pneumoniae and Plays a Protective Role in Pneumococcal Pneumonia
      PLoS Pathog, 2016;12(12):e1006038.
      Species: Mouse
      Sample Type: Tissue Homogenates
    4. Prion infection of mouse brain reveals multiple new upregulated genes involved in neuroinflammation or signal transduction.
      Authors: Carroll J, Striebel J, Race B, Phillips K, Chesebro B
      J Virol, 2015;89(4):2388-404.
      Species: Mouse
      Sample Type: Tissue Homogenates
    5. Sharpin Controls Osteogenic Differentiation of Mesenchymal Bone Marrow Cells.
      Authors: Jeschke A, Catala-Lehnen P, Sieber S, Bickert T, Schweizer M, Koehne T, Wintges K, Marshall R, Mautner A, Duchstein L, Otto B, Horst A, Amling M, Kreienkamp H, Schinke T
      J Immunol, 2015;195(8):3675-84.
      Species: Mouse
      Sample Type: Serum
    6. Correction: Yersinia pestis Activates Both IL-1beta and IL-1 Receptor Antagonist to Modulate Lung Inflammation during Pneumonic Plague.
      PLoS Pathog, 2015;11(5):e1004921.
      Species: Mouse
      Sample Type: BALF
    7. Inflammatory bone loss in experimental periodontitis induced by Aggregatibacter actinomycetemcomitans in interleukin-1 receptor antagonist knockout mice.
      Authors: Izawa A, Ishihara Y, Mizutani H, Kobayashi S, Goto H, Okabe E, Takeda H, Ozawa Y, Kamiya Y, Sugita Y, Kubo K, Kamei H, Kikuchi T, Mitani A, Hayashi J, Nishihara T, Maeda H, Noguchi T
      Infect Immun, 2014;82(5):1904-13.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    8. The multifaceted effects of polysaccharides isolated from Dendrobium huoshanense on immune functions with the induction of interleukin-1 receptor antagonist (IL-1ra) in monocytes.
      Authors: Lin J, Chang Y, Yang W, Yu A, Wong C
      PLoS ONE, 2014;9(4):e94040.
      Species: Mouse
      Sample Type: Serum
    9. Inflammation induced by MMP-9 enhances tumor regression of experimental breast cancer.
      Authors: Leifler K, Svensson S, Abrahamsson A, Bendrik C, Robertson J, Gauldie J, Olsson A, Dabrosin C
      J Immunol, 2013;190(8):4420-30.
      Species: Mouse
      Sample Type: Tissue Homogenates
    10. Impaired bone formation and increased osteoclastogenesis in mice lacking chemokine (C-C motif) ligand 5 (Ccl5).
      Authors: Wintges K, Beil F, Albers J, Jeschke A, Schweizer M, Claass B, Tiegs G, Amling M, Schinke T
      J Bone Miner Res, 2013;28(10):2070-80.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    11. Human mesenchymal stem/stromal cells cultured as spheroids are self-activated to produce prostaglandin E2 that directs stimulated macrophages into an anti-inflammatory phenotype.
      Authors: Ylostalo J, Bartosh T, Coble K, Prockop D
      Stem Cells, 2012;30(10):2283-96.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    12. Fasting induces an anti-inflammatory effect on the neuroimmune system which a high-fat diet prevents.
      Authors: Lavin DN, Joesting JJ, Chiu GS, Moon ML, Meng J, Dilger RN, Freund GG
      Obesity (Silver Spring), 2011;19(8):1586-94.
      Species: Mouse
      Sample Type: Serum
    13. TLR4 is a negative regulator in noninfectious lung inflammation.
      Authors: Zhao H, Leu SW, Shi L, Dedaj R, Zhao G, Garg HG, Shen L, Lien E, Fitzgerald KA, Shiedlin A, Shen H, Quinn DA, Hales CA
      J. Immunol., 2010;184(9):5308-14.
      Species: Mouse
      Sample Type: BALF
    14. Activation of NOD2 in vivo induces IL-1beta production in the eye via caspase-1 but results in ocular inflammation independently of IL-1 signaling.
      Authors: Rosenzweig HL, Martin TM, Planck SR, Galster K, Jann MM, Davey MP, Kobayashi K, Flavell RA, Rosenbaum JT
      J. Leukoc. Biol., 2008;84(2):529-36.
      Species: Mouse
      Sample Type: Tissue Homogenates
    15. A role for IL-1 receptor antagonist or other cytokines in the acute therapeutic effects of IVIg?
      Authors: Crow AR, Song S, Semple JW, Freedman J, Lazarus AH
      Blood, 2007;109(1):155-8.
      Species: Mouse
      Sample Type: Serum
    16. Interleukin 1 receptor antagonist mediates the antiinflammatory and antifibrotic effect of mesenchymal stem cells during lung injury.
      Authors: Ortiz LA, Dutreil M, Fattman C, Pandey AC, Torres G, Go K, Phinney DG
      Proc. Natl. Acad. Sci. U.S.A., 2007;104(26):11002-7.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    17. Intrapulmonary delivery of bone marrow-derived mesenchymal stem cells improves survival and attenuates endotoxin-induced acute lung injury in mice.
      Authors: Gupta N, Su X, Popov B, Lee JW, Serikov V, Matthay MA
      J. Immunol., 2007;179(3):1855-63.
      Species: Mouse
      Sample Type: BALF
    18. Absence of IL-1 receptor antagonist impaired wound healing along with aberrant NF-kappaB activation and a reciprocal suppression of TGF-beta signal pathway.
      Authors: Ishida Y, Kondo T, Kimura A, Matsushima K, Mukaida N
      J. Immunol., 2006;176(9):5598-606.
      Species: Mouse
      Sample Type: Tissue Homogenates
    19. Interleukin-1 beta contributes to the upregulation of kappa opioid receptor mrna in dorsal root ganglia in response to peripheral inflammation.
      Authors: Puehler W, Rittner HL, Mousa SA, Brack A, Krause H, Stein C, Schafer M
      Neuroscience, 2006;141(2):989-98.
      Species: Rat
      Sample Type: Cell Culture Supernates
    20. A deficit in astroglial organization causes the impaired reactive sprouting in human apolipoprotein E4 targeted replacement mice.
      Authors: Blain JF, Sullivan PM, Poirier J
      Neurobiol. Dis., 2005;21(3):505-14.
      Species: Mouse
      Sample Type: Tissue Homogenates
    21. IL-1 receptor-associated kinase 1 mediates protection against Staphylococcus aureus infection.
      Authors: Verdrengh M, Thomas JA, Hultgren OH
      Microbes Infect., 2004;6(14):1268-72.
      Species: Mouse
      Sample Type: Serum
    22. Regulation of Pseudomonas aeruginosa corneal infection in IL-1 beta converting enzyme (ICE, caspase-1) deficient mice.
      Authors: Thakur A, Barrett RP, McClellan S, Hazlett LD
      Curr. Eye Res., 2004;29(4):225-33.
      Species: Mouse
      Sample Type: Tissue Homogenates
    Expand to show all 22 Citations
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