Mouse IL-27 p28/IL-30 Quantikine ELISA Kit

(13 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
    4.7 pg/mL
  • Assay Range
    7.8 - 500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant mouse IL-27 p28
  • Cross-reactivity
    Cross-reactivity observed with 1 or more available related molecules.Cross-species reactivity not tested.
  • Interference
    Interference observed with 1 or more available related molecules.
Product Summary
The Quantikine Mouse IL-27 p28 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse IL-27 p28 in cell culture supernates, mouse serum, and plasma. It contains NS0-expressed recombinant mouse IL-27 p28 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse IL-27 p28 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-27 p28.

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, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Standard Deviation1.83.414.62.34.521.3


The recovery of mouse IL-27 p28 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=8) 102 83-112
EDTA Plasma (n=8) 99 83-116
Heparin Plasma (n=8) 95 87-101
Serum (n=8) 95 85-102
To assess the linearity of the assay, samples containing or spiked with high concentrations of mouse IL-27 p28 in each matrix were diluted with Calibrator Diluent and then assayed.
Mouse IL-27 p28/IL-30 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: IL-27 p28/IL-30

Interleukin-27 (IL-27) is a heterodimeric group 2 receptor ligand molecule that belongs to the IL-6/IL-12 family of long type I cytokines. It is composed of EBI3 (EBV-induced gene 3), a 34 kDa glycoprotein that is related to the p40 subunit of IL-12 and IL-23, and p28, the 28 kDa glycoprotein that is related to the p35 chain of IL-12). The human EBI3 gene encodes a 229 amino acid (aa) precursor that contains a 20 aa signal peptide and 209 aa mature protein. The mature region contains two potential N-linked glycosylation sites, two fibronectin type III domains, and two pairs of conserved cysteine residues with a WSxWS-like motif that places the IL-27 molecule in the hematopoietin receptor family. Although p40, the EBI3 counterpart in IL-12, is known to form homodimers, there is no evidence to date that EBI3 also homodimerizes.

Human EBI3 is 61% aa identical to mouse EBI3. The human p28 gene encodes a 243 aa precursor that contains a 28 aa signal sequence and 215 aa mature region. The mature region is characterized by the presence of four α-helices, placing it in the IL-6 family of helical cytokines. Human p28 is 74% aa identical to mouse p28. IL-27 is expressed by monocytes, endothelial cells and dendritic cells. IL-27 binds to and signals through a heterodimeric receptor complex composed of WSX-1 (TCCR) and gp130. Evidence suggests IL-27 interacts only with WSX-1. IL-27 has both anti- and pro-inflammatory properties. As an anti-inflammatory, IL-27 seems to induce a general negative feedback program that limits T and NK-T cell activity. At the onset of infection, IL-27 induces an IL-12 receptor on naïve CD4+ T cells, making them susceptible to subsequent IL-12 activity (and possible Th1 development).

    • Long Name
      Interleukin 27 p28
    • Entrez Gene IDs
      246778 (Human); 246779 (Mouse);
    • Alternate Names
      IL27 p28; IL-30;
    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.

    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.

    Showing Results 1 - 10 of 13
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    Sample Type
    1. Interleukin-35 Inhibits Endothelial Cell Activation by Suppressing MAPK-AP-1 Pathway.
      Authors: Sha X, Meng S, Li X, Xi H, Maddaloni M, Pascual D, Shan H, Jiang X, Wang H, Yang X
      J Biol Chem, 2015;290(31):19307-18.
      Species: Mouse
      Sample Type: Plasma
    2. IL-27R-mediated regulation of IL-17 controls the development of respiratory syncytial virus-associated pathogenesis.
      Authors: de Almeida Nagata D, Demoor T, Ptaschinski C, Ting H, Jang S, Reed M, Mukherjee S, Lukacs N
      Am J Pathol, 2014;184(6):1807-18.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    3. IL-10-Producing Th1 Cells and Disease Progression Are Regulated by Distinct CD11c+ Cell Populations during Visceral Leishmaniasis.
      Authors: Owens BM, Beattie L, Moore JW, Brown N, Mann JL, Dalton JE, Maroof A, Kaye PM
      PLoS Pathog., 2012;8(7):e1002827.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    4. A role for IL-27p28 as an antagonist of gp130-mediated signaling.
      Authors: Stumhofer JS, Tait ED, Quinn WJ, Hosken N, Spudy B, Goenka R, Fielding CA, O'Hara AC, Chen Y, Jones ML, Saris CJ, Rose-John S, Cua DJ, Jones SA, Elloso MM, Grotzinger J, Cancro MP, Levin SD, Hunter CA
      Nat. Immunol., 2010;11(12):1119-26.
      Species: Mouse
      Sample Type: Serum
    5. Critical role of the IFN-stimulated gene factor 3 complex in TLR-mediated IL-27p28 gene expression revealing a two-step activation process.
      Authors: Molle C, Goldman M, Goriely S
      J. Immunol., 2010;184(4):1784-92.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    6. Modified adenine (9-benzyl-2-butoxy-8-hydroxyadenine) redirects Th2-mediated murine lung inflammation by triggering TLR7.
      Authors: Vultaggio A, Nencini F, Fitch PM, Fili L, Maggi L, Fanti P, deVries A, Beccastrini E, Palandri F, Manuelli C, Bani D, Giudizi MG, Guarna A, Annunziato F, Romagnani S, Maggi E, Howie SE, Parronchi P
      J. Immunol., 2009;182(2):880-9.
      Species: Mouse
      Sample Type: Serum
    7. Engagement of the type I interferon receptor on dendritic cells inhibits T helper 17 cell development: role of intracellular osteopontin.
      Authors: Shinohara ML, Kim JH, Garcia VA, Cantor H
      Immunity, 2008;29(1):68-78.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    8. The type I IFN induction pathway constrains Th17-mediated autoimmune inflammation in mice.
      Authors: Guo B, Chang EY, Cheng G
      J. Clin. Invest., 2008;118(5):1680-90.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    9. Peroxisome proliferator-activated receptor-alpha agonist fenofibrate regulates IL-12 family cytokine expression in the CNS: relevance to multiple sclerosis.
      Authors: Xu J, Racke MK, Drew PD
      J. Neurochem., 2007;103(5):1801-10.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    10. Regulation of IL-27 p28 gene expression in macrophages through MyD88- and interferon-gamma-mediated pathways.
      Authors: Liu J, Guan X, Ma X
      J. Exp. Med., 2007;204(1):141-52.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    11. IL-27 synthesis induced by TLR ligation critically depends on IFN regulatory factor 3.
      Authors: Molle C, Nguyen M, Flamand V, Renneson J, Trottein F, De Wit D, Willems F, Goldman M, Goriely S
      J. Immunol., 2007;178(12):7607-15.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    12. The proinflammatory effect of prostaglandin E2 in experimental inflammatory bowel disease is mediated through the IL-23-->IL-17 axis.
      Authors: Sheibanie AF, Yen JH, Khayrullina T, Emig F, Zhang M, Tuma R, Ganea D
      J. Immunol., 2007;178(12):8138-47.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    13. Suppressive effect of IL-27 on encephalitogenic Th17 cells and the effector phase of experimental autoimmune encephalomyelitis.
      Authors: Fitzgerald DC, Ciric B, Touil T, Harle H, Grammatikopolou J, Das Sarma J, Gran B, Zhang GX, Rostami A
      J. Immunol., 2007;179(5):3268-75.
      Species: Mouse
      Sample Type: Cell Culture Supernates
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