Mouse IL-12 p40 Allele-specific Quantikine ELISA Kt

(27 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)
  • Sensitivity
    4 pg/mL
  • Assay Range
    11.7 - 750 pg/mL (Cell Culture Supernates, Serum)
  • 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 and mouse serum. It contains recombinant mouse IL-12 p40 homodimer and antibodies raised against the recombinant factor. 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
Intra-Assay Precision Inter-Assay Precision
Standard Deviation0.811.


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

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=11) 100 81-116
Serum (n=7) 104 85-115
To assess the linearity of the assay, five 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 Allele-specific Quantikine ELISA Kt
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 27
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    Sample Type
    1. The adaptor molecule CARD9 is essential for tuberculosis control.
      Authors: Dorhoi A, Desel C, Yeremeev V, Pradl L, Brinkmann V, Mollenkopf HJ, Hanke K, Gross O, Ruland J, Kaufmann SH
      J. Exp. Med., 2011;207(4):777-92.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    2. Proinflammatory clearance of apoptotic neutrophils induces an IL-12(low)IL-10(high) regulatory phenotype in macrophages.
      Authors: Filardy AA, Pires DR, Nunes MP, Takiya CM, Freire-de-Lima CG, Ribeiro-Gomes FL, DosReis GA
      J. Immunol., 2010;185(4):2044-50.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    3. NOD1 contributes to mouse host defense against Helicobacter pylori via induction of type I IFN and activation of the ISGF3 signaling pathway.
      Authors: Watanabe T, Asano N, Fichtner-Feigl S, Gorelick PL, Tsuji Y, Matsumoto Y, Chiba T, Fuss IJ, Kitani A, Strober W
      J. Clin. Invest., 2010;120(5):1645-62.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    4. Activation of the NLRP3 inflammasome in dendritic cells induces IL-1beta-dependent adaptive immunity against tumors.
      Authors: Ghiringhelli F, Apetoh L, Tesniere A, Aymeric L, Ma Y, Ortiz C, Vermaelen K, Panaretakis T, Mignot G, Ullrich E, Perfettini JL, Schlemmer F, Tasdemir E, Uhl M, Genin P, Civas A, Ryffel B, Kanellopoulos J, Tschopp J, Andre F, Lidereau R, McLaughlin NM, Haynes NM, Smyth MJ, Kroemer G, Zitvogel L
      Nat. Med., 2009;15(10):1170-8.
      Species: Mouse
      Sample Type: Cells Supernates
    5. 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
    6. An eosinophil immune response characterizes the inflammatory skin disease observed in Tie-2 transgenic mice.
      Authors: Voskas D, Babichev Y, Ling LS, Alami J, Shaked Y, Kerbel RS, Ciruna B, Dumont DJ
      J. Leukoc. Biol., 2008;84(1):59-67.
      Species: Mouse
      Sample Type: Serum
    7. 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
    8. TLR-2 and IL-17A in chitin-induced macrophage activation and acute inflammation.
      Authors: Da Silva CA, Hartl D, Liu W, Lee CG, Elias JA
      J. Immunol., 2008;181(6):4279-86.
      Species: Mouse
      Sample Type: BALF
    9. Morphine withdrawal inhibits IL-12 induction in a macrophage cell line through a mechanism that involves cAMP.
      Authors: Kelschenbach J, Ninkovic J, Wang J, Krishnan A, Charboneau R, Barke RA, Roy S
      J. Immunol., 2008;180(6):3670-9.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    10. Bcl-xL antisense oligonucleotide and cisplatin combination therapy extends survival in SCID mice with established mesothelioma xenografts.
      Authors: Littlejohn JE, Cao X, Miller SD, Ozvaran MK, Jupiter D, Zhang L, Rodarte C, Smythe WR
      Int. J. Cancer, 2008;123(1):202-8.
      Species: Mouse
      Sample Type: Serum
    11. The direct action of 1alpha,25(OH)2-vitamin D3 on purified mouse Langerhans cells.
      Authors: Fujita H, Asahina A, Komine M, Tamaki K
      Cell. Immunol., 2007;245(2):70-9.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    12. Intercellular adhesion molecule-1 expression is required on multiple cell types for the development of experimental autoimmune encephalomyelitis.
      Authors: Bullard DC, Hu X, Schoeb TR, Collins RG, Beaudet AL, Barnum SR
      J. Immunol., 2007;178(2):851-7.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    13. Inactivated Sendai virus particles eradicate tumors by inducing immune responses through blocking regulatory T cells.
      Authors: Kurooka M, Kaneda Y
      Cancer Res., 2007;67(1):227-36.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    14. Toll-like receptor 4 mediates the early inflammatory response after cold ischemia/reperfusion.
      Authors: Kaczorowski DJ, Nakao A, Mollen KP, Vallabhaneni R, Sugimoto R, Kohmoto J, Tobita K, Zuckerbraun BS, McCurry KR, Murase N, Billiar TR
      Transplantation, 2007;84(10):1279-87.
      Species: Mouse
      Sample Type: Serum
    15. Mrp8 and Mrp14 are endogenous activators of Toll-like receptor 4, promoting lethal, endotoxin-induced shock.
      Authors: Vogl T, Tenbrock K, Ludwig S, Leukert N, Ehrhardt C, van Zoelen MA, Nacken W, Foell D, van der Poll T, Sorg C, Roth J
      Nat. Med., 2007;13(9):1042-9.
      Species: Mouse
      Sample Type: Serum
    16. Vaccinia virus infection attenuates innate immune responses and antigen presentation by epidermal dendritic cells.
      Authors: Deng L, Dai P, Ding W, Granstein RD, Shuman S
      J. Virol., 2006;80(20):9977-87.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    17. Induction of dendritic cell maturation by pertussis toxin and its B subunit differentially initiate Toll-like receptor 4-dependent signal transduction pathways.
      Authors: Wang ZY, Yang D, Chen Q, Leifer CA, Segal DM, Su SB, Caspi RR, Howard ZO, Oppenheim JJ
      Exp. Hematol., 2006;34(8):1115-24.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    18. Roles of the small intestine for induction of toll-like receptor 4-mediated innate resistance in naturally acquired murine toxoplasmosis.
      Authors: Furuta T, Yoshikawa Y
      Int. Immunol., 2006;18(12):1655-62.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    19. A multivalent combination of experimental antituberculosis DNA vaccines based on Ag85B and regions of difference antigens.
      Authors: Grover A, Ahmed MF, Singh B, Verma I, Sharma P, Khuller GK
      Microbes Infect., 2006;8(9):2390-9.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    20. Endogenous interleukin-12 improves the early antimicrobial host response to murine Escherichia coli peritonitis.
      Authors: Weijer S, Florquin S, van der Poll T
      Shock, 2005;23(1):54-8.
      Species: Mouse
      Sample Type: Peritoneal Fluid
    21. Cutting edge: Physiologic attenuation of proinflammatory transcription by the Gs protein-coupled A2A adenosine receptor in vivo.
      Authors: Lukashev D, Ohta A, Apasov S, Chen JF, Sitkovsky M
      J. Immunol., 2004;173(1):21-4.
      Species: Mouse
      Sample Type: Peritoneal Exudate
    22. Chemokine and cytokine production during Orientia tsutsugamushi infection in mice.
      Authors: Koh YS, Yun JH, Seong SY, Choi MS, Kim IS
      Microb. Pathog., 2004;36(1):51-7.
      Species: Mouse
      Sample Type: Peritoneal Fluid
    23. IL-12 p40 homodimer-dependent macrophage chemotaxis and respiratory viral inflammation are mediated through IL-12 receptor beta 1.
      Authors: Russell TD, Yan Q, Fan G, Khalifah AP, Bishop DK, Brody SL, Walter MJ
      J. Immunol., 2003;171(12):6866-74.
      Species: Mouse
      Sample Type: BALF
    24. A profibrotic function of IL-12p40 in experimental pulmonary fibrosis.
      Authors: Huaux F, Arras M, Tomasi D, Barbarin V, Delos M, Coutelier JP, Vink A, Phan SH, Renauld JC, Lison D
      J. Immunol., 2002;169(5):2653-61.
      Species: Mouse
      Sample Type: Tissue Homogenates
    25. Administration of either anti-CD40 or interleukin-12 following lethal total body irradiation induces acute lethal toxicity affecting the gut.
      Authors: Hixon JA, Anver MR, Blazar BR, Panoskaltsis-Mortari A, Wiltrout RH, Murphy WJ
      Biol. Blood Marrow Transplant., 2002;8(6):316-25.
      Species: Mouse
      Sample Type: Serum
    26. 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
    27. In vitro and in vivo immunostimulatory potential of bone marrow-derived mast cells on B- and T-lymphocyte activation.
      Authors: Tkaczyk C, Villa I, Peronet R, David B, Chouaib S, Mecheri S
      J. Allergy Clin. Immunol., 2000;105(1):134-42.
      Species: Mouse
      Sample Type: Cell Culture Supernates
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