Human IL-12/IL-23 p40 Quantikine ELISA Kit

(21 citations)   
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    3.5 hours or 4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (100 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL), Citrate Plasma (100 uL)
  • Sensitivity
    15 pg/mL
  • Assay Range
    31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
  • Specificity
    Natural and recombinant human IL-12 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.
Control Available
Product Summary
The Quantikine Human IL-12 p40 Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure IL-12 p40 in cell culture supernates, serum, and plasma. It contains recombinant human IL-12 p40 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate recombinant human IL-12 p40. Results obtained using natural human IL-12 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 human IL-12 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.
Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma
Intra-Assay Precision Inter-Assay Precision
Standard Deviation9.927.262.31122.363

Cell Culture Supernates
Intra-Assay Precision Inter-Assay Precision
Standard Deviation5.111.435.511.819.860.6


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

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 96 85-105
Citrate Plasma (n=5) 100 85-112
EDTA Plasma (n=5) 101 92-113
Heparin Plasma (n=5) 96 89-106
Serum (n=5) 98 86-112
To assess the linearity of the assay, five samples were spiked with high concentrations of IL-12 p40 in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human IL-12/IL-23 p40 Quantikine ELISA Kit
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. 100 µL Assay Diluent
    4.   Add 100 µL of Assay Diluent to each well.

    5. 100 µL Standard, Control, or Sample
    6.   Add 100 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 3 times for a total of 4 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well.
    10.   For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
      For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour.
    11.   Aspirate and wash 4 times.

    12. 200 µL Substrate Solution
    13.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

    14. 50 µL Stop Solution
    15. Add 50 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 21
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    Sample Type
    1. Dysregulated serum IL-23 and SIRT1 activity in peripheral blood mononuclear cells of patients with rheumatoid arthritis.
      Authors: Wendling D, Abbas W, Godfrin-Valnet M, Kumar A, Guillot X, Khan K, Vidon C, Coquard L, Toussirot E, Prati C, Herbein G
      PLoS ONE, 2015;10(3):e0119981.
      Species: Human
      Sample Type: Serum
    2. Free IL-12p40 monomer is a polyfunctional adaptor for generating novel IL-12-like heterodimers extracellularly.
      Authors: Abdi K, Singh N, Spooner E, Kessler B, Radaev S, Lantz L, Xiao T, Matzinger P, Sun P, Ploegh H
      J Immunol, 2014;192(12):6028-36.
      Species: Mouse
      Sample Type: Cell culture supernatant
    3. Co-culture of human bronchial fibroblasts and CD4+ T cells increases Th17 cytokine signature.
      Authors: Loubaki L, Hadj-Salem I, Fakhfakh R, Jacques E, Plante S, Boisvert M, Aoudjit F, Chakir J
      PLoS ONE, 2013;8(12):e81983.
      Species: Human
      Sample Type: cell culture supernate
    4. Direct and indirect impairment of human dendritic cell function by virulent Francisella tularensis Schu S4.
      Authors: Chase JC, Celli J, Bosio CM
      Infect. Immun., 2009;77(1):180-95.
      Species: Human
      Sample Type: Cell Culture Supernates
    5. Evidence for a pathogenetic role of interleukin-18 in cutaneous lupus erythematosus.
      Authors: Wang D, Drenker M, Eiz-Vesper B, Werfel T, Wittmann M
      Arthritis Rheum., 2008;58(10):3205-15.
      Species: Human
      Sample Type: Cell Culture Supernates
    6. Distinct subsets of human invariant NKT cells differentially regulate T helper responses via dendritic cells.
      Authors: Liu TY, Uemura Y, Suzuki M, Narita Y, Hirata S, Ohyama H, Ishihara O, Matsushita S
      Eur. J. Immunol., 2008;38(4):1012-23.
      Species: Human
      Sample Type: Cell Culture Supernates
    7. The 3'UTR 1188 A/C polymorphism in the interleukin-12p40 gene (IL-12B) is associated with lepromatous leprosy in the west of Mexico.
      Authors: Alvarado-Navarro A, Montoya-Buelna M, Munoz-Valle JF, Lopez-Roa RI, Guillen-Vargas C, Fafutis-Morris M
      Immunol. Lett., 2008;118(2):148-51.
      Species: Human
      Sample Type: Serum
    8. 1, 25 Dihydroxyvitamin D3 modulated cytokine response in pulmonary tuberculosis.
      Authors: Vidyarani M, Selvaraj P, Jawahar MS, Narayanan PR
      Cytokine, 2007;40(2):128-34.
      Species: Human
      Sample Type: Cell Culture Supernates
    9. Chemokines CXCL10 and CXCL11 in the cerebrospinal fluid of patients with tick-borne encephalitis.
      Authors: Lepej SZ, Misic-Majerus L, Jeren T, Rode OD, Remenar A, Sporec V, Vince A
      Acta Neurol. Scand., 2007;115(2):109-14.
      Species: Human
      Sample Type: Serum
    10. Placental growth factor down-regulates type 1 T helper immune response by modulating the function of dendritic cells.
      Authors: Lin YL, Liang YC, Chiang BL
      J. Leukoc. Biol., 2007;82(6):1473-80.
      Species: Human
      Sample Type: Cell Culture Supernates
    11. Impairment of human NK cell cytotoxic activity and cytokine release by cigarette smoke.
      Authors: Mian MF, Lauzon NM, Stampfli MR, Mossman KL, Ashkar AA
      J. Leukoc. Biol., 2007;83(3):774-84.
      Species: Human
      Sample Type: Cell Culture Supernates
    12. First in human phase I trial of 852A, a novel systemic toll-like receptor 7 agonist, to activate innate immune responses in patients with advanced cancer.
      Authors: Dudek AZ, Yunis C, Harrison LI, Kumar S, Hawkinson R, Cooley S, Vasilakos JP, Gorski KS, Miller JS
      Clin. Cancer Res., 2007;13(23):7119-25.
      Species: Human
      Sample Type: Serum
    13. Oral resiquimod in chronic HCV infection: safety and efficacy in 2 placebo-controlled, double-blind phase IIa studies.
      Authors: Pockros PJ, Guyader D, Patton H, Tong MJ, Wright T, McHutchison JG, Meng TC
      J. Hepatol., 2007;47(2):174-82.
      Species: Human
      Sample Type: Serum
    14. IL-23 is increased in dendritic cells in multiple sclerosis and down-regulation of IL-23 by antisense oligos increases dendritic cell IL-10 production.
      Authors: Vaknin-Dembinsky A, Balashov K, Weiner HL
      J. Immunol., 2006;176(12):7768-74.
      Species: Human
      Sample Type: Cell Culture Supernates
    15. Bordetella pertussis inhibition of interleukin-12 (IL-12) p70 in human monocyte-derived dendritic cells blocks IL-12 p35 through adenylate cyclase toxin-dependent cyclic AMP induction.
      Authors: Spensieri F, Fedele G, Fazio C, Nasso M, Stefanelli P, Mastrantonio P, Ausiello CM
      Infect. Immun., 2006;74(5):2831-8.
      Species: Human
      Sample Type: Cell Culture Supernates
    16. Elevation of IL-12 p40 and its antibody in myasthenia gravis with thymoma.
      Authors: Yoshikawa H, Sato K, Edahiro S, Furukawa Y, Maruta T, Iwasa K, Watanabe H, Takaoka S, Suzuki Y, Takamori M, Yamada M
      J. Neuroimmunol., 2006;175(1):169-75.
      Species: Human
      Sample Type: Serum
    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: Human
      Sample Type: Cell Culture Supernates
    18. Human complete Stat-1 deficiency is associated with defective type I and II IFN responses in vitro but immunity to some low virulence viruses in vivo.
      Authors: Chapgier A, Wynn RF, Jouanguy E, Filipe-Santos O, Zhang S, Feinberg J, Hawkins K, Casanova JL, Arkwright PD
      J. Immunol., 2006;176(8):5078-83.
      Species: Human
      Sample Type: Plasma
    19. Experimental heatstroke in baboon: analysis of the systemic inflammatory response.
      Authors: Bouchama A, Ollivier V, Roberts G, Al Mohanna F, de Prost D, Eldali A, Saussereau E, El-Sayed R, Chollet-Martin S
      Shock, 2005;24(4):332-5.
      Species: Primate – Papio hamadryas (Hamadyras Baboon)
      Sample Type: Plasma
    20. Cytokines and adhesion molecules in multiple sclerosis patients treated with interferon-beta1b.
      Authors: Jensen J, Krakauer M, Sellebjerg F
      Cytokine, 2005;29(1):24-30.
      Species: Human
      Sample Type: Plasma
    21. HIV-1 protein Vpr suppresses IL-12 production from human monocytes by enhancing glucocorticoid action: potential implications of Vpr coactivator activity for the innate and cellular immunity deficits observed in HIV-1 infection.
      Authors: Mirani M, Elenkov I, Volpi S, Hiroi N, Chrousos GP, Kino T
      J. Immunol., 2002;169(11):6361-8.
      Species: Human
      Sample Type: Cell Culture Supernates
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