Mouse IL-12 p40 NonAllele-specific Quantikine ELISA

(16 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 (25 uL), EDTA Plasma (25 uL), Heparin Plasma (25 uL)
  • Sensitivity
    2.7 pg/mL
  • Assay Range
    11.7 - 750 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant mouse IL-12/IL-23 p40
  • Cross-reactivity
    Cross-reactivity observed with 1 or more 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 IL-12/IL-23 p40 Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse p40 subunit in IL-12 p70, IL-23, p40 homodimer, and p40 monomer in cell culture supernates, mouse serum, and plasma. It contains recombinant mouse IL-12/23 p40 homodimer and antibodies raised against the p40 subunit of recombinant mouse IL-12 p70. This immunoassay has been shown to accurately quantitate the recombinant mouse IL-12/IL-23 p40. Results obtained using natural mouse IL-12/IL-23 p40 showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse IL-12/IL-23 p40.

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
Standard Deviation2.3317.23.86.533.7


The recovery of mouse IL-12/IL-23 p40 spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=4) 104 98-108
EDTA Plasma (n=4) 99 81-112
Heparin Plasma (n=4) 95 80-109
Serum (n=4) 105 86-117
To assess the linearity of the assay, four or more samples containing and/or spiked with various concentrations of mouse IL-12/IL-23 p40 in each matrix were diluted with Calibrator Diluent and then assayed.
Mouse IL-12 p40 NonAllele-specific Quantikine ELISA
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: IL-12/IL-23 p40

Interleukin 12 (IL-12), also known as natural killer cell stimulatory factor (NKSF) or cytotoxic lymphocyte maturation factor (CLMF), is a heterodimeric pleiotropic cytokine made up of a 40 kDa (p40) subunit and a 35 kDa (p35) subunit. The IL-12 p40 subunit is shared by IL-23, another heterodimeric cytokine that has biological activities similar to, as well as distinct from, IL-12. IL-12 is produced by macrophages and B cells and has been shown to have multiple effects on T cells and natural killer (NK) cells. While mouse IL-12 is active on both human and mouse cells, human IL-12 is not active on mouse cells.

    • Long Name
      Interleukin 12/Interleukin 23 p40
    • Entrez Gene IDs
      3593 (Human); 16160 (Mouse); 64546 (Rat); 403976 (Canine); 493741 (Feline);
    • Alternate Names
      CLMF2; IL12 p40; IL12B; IL-12B; NKSF2;
    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 16
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    Sample Type
    1. 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
    2. Modelling of mouse experimental colitis by global property screens: a holistic approach to assess drug effects in inflammatory bowel disease.
      Authors: Gottfries J, Melgar S, Michaelsson E
      PLoS ONE, 2012;7(1):e30005.
      Species: Mouse
      Sample Type: Tissue Homogenates
    3. Cutting edge: Divergent cell-specific functions of MyD88 for inflammatory responses and organ injury in septic peritonitis.
      Authors: Gais P, Reim D, Jusek G, Rossmann-Bloeck T, Weighardt H, Pfeffer K, Altmayr F, Janssen KP, Holzmann B
      J. Immunol., 2012;188(12):5833-7.
      Species: Mouse
      Sample Type: Plasma
    4. ChemR23 dampens lung inflammation and enhances anti-viral immunity in a mouse model of acute viral pneumonia.
      Authors: Bondue B, Vosters O, de Nadai P, Glineur S, De Henau O, Luangsay S, Van Gool F, Communi D, De Vuyst P, Desmecht D, Parmentier M
      PLoS Pathog., 2011;7(11):e1002358.
      Species: Mouse
      Sample Type: Tissue Homogenates
    5. Serotonin activates dendritic cell function in the context of gut inflammation.
      Authors: Li N, Ghia JE, Wang H, McClemens J, Cote F, Suehiro Y, Mallet J, Khan WI
      Am. J. Pathol., 2011;178(2):662-71.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    6. p47phox deficiency induces macrophage dysfunction resulting in progressive crystalline macrophage pneumonia.
      Authors: Liu Q, Cheng LI, Yi L, Zhu N, Wood A, Changpriroa CM, Ward JM, Jackson SH
      Am. J. Pathol., 2009;174(1):153-63.
      Species: Mouse
      Sample Type: BALF
    7. Unc93B1 biases Toll-like receptor responses to nucleic acid in dendritic cells toward DNA- but against RNA-sensing.
      Authors: Fukui R, Saitoh S, Matsumoto F, Kozuka-Hata H, Oyama M, Tabeta K, Beutler B, Miyake K
      J. Exp. Med., 2009;206(6):1339-50.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    8. NOD2-deficient mice have impaired resistance to Mycobacterium tuberculosis infection through defective innate and adaptive immunity.
      Authors: Divangahi M, Mostowy S, Coulombe F, Kozak R, Guillot L, Veyrier F, Kobayashi KS, Flavell RA, Gros P, Behr MA
      J. Immunol., 2008;181(10):7157-65.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    9. Cytokine production by M-CSF- and GM-CSF-induced mouse bone marrow-derived macrophages upon coculturing with late apoptotic cells.
      Authors: Yamazaki T, Nagata K, Kobayashi Y
      Cell. Immunol., 2008;251(2):124-30.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    10. Cigarette smoke selectively enhances viral PAMP- and virus-induced pulmonary innate immune and remodeling responses in mice.
      Authors: Kang MJ, Lee CG, Lee JY, Dela Cruz CS, Chen ZJ, Enelow R, Elias JA
      J. Clin. Invest., 2008;118(8):2771-84.
      Species: Mouse
      Sample Type: BALF
    11. Pulmonary stromal cells induce the generation of regulatory DC attenuating T-cell-mediated lung inflammation.
      Authors: Li Q, Guo Z, Xu X, Xia S, Cao X
      Eur. J. Immunol., 2008;38(10):2751-61.
      Species: Mouse
      Sample Type: BALF
    12. Toll-like receptor (TLR)2 and TLR3 synergy and cross-inhibition in murine myeloid dendritic cells.
      Authors: Vanhoutte F, Paget C, Breuilh L, Fontaine J, Vendeville C, Goriely S, Ryffel B, Faveeuw C, Trottein F
      Immunol. Lett., 2007;116(1):86-94.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    13. Oral administration of an immunostimulatory DNA sequence from Bifidobacterium longum improves Th1/Th2 balance in a murine model.
      Authors: Takahashi N, Kitazawa H, Iwabuchi N, Xiao JZ, Miyaji K, Iwatsuki K, Saito T
      Biosci. Biotechnol. Biochem., 2006;70(8):2013-7.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    14. A novel siRNA-lipoplex technology for RNA interference in the mouse vascular endothelium.
      Authors: Santel A, Aleku M, Keil O, Endruschat J, Esche V, Fisch G, Dames S, Loffler K, Fechtner M, Arnold W, Giese K, Klippel A, Kaufmann J
      Gene Ther., 2006;13(16):1222-34.
      Species: Mouse
      Sample Type: Serum
    15. The transcriptional response to lipopolysaccharide reveals a role for interferon-gamma in lung neutrophil recruitment.
      Authors: Burch LH, Yang IV, Whitehead GS, Chao FG, Berman KG, Schwartz DA
      Am. J. Physiol. Lung Cell Mol. Physiol., 2006;291(4):L677-82.
      Species: Mouse
      Sample Type: BALF
    16. Dendritic cells phagocytose and are activated by Treponema pallidum.
      Authors: Bouis DA, Popova TG, Takashima A, Norgard MV
      Infect. Immun., 2001;69(1):518-28.
      Species: Mouse
      Sample Type: Cell Culture Supernates
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