Human IL-12 p70 Quantikine ELISA Kit

(44 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 (200 uL), Serum (200 uL), EDTA Plasma (200 uL), Heparin Plasma (200 uL), Citrate Plasma (200 uL), Urine (200 uL)
  • Sensitivity
    5 pg/mL
  • Assay Range
    7.8 - 500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine)
  • Specificity
    Natural and recombinant human IL-12p70
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC01-1, Quantikine Immunoassay Control Group 1 - Please Inquire
Product Summary
The Quantikine Human IL-12 Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure IL-12 in cell culture supernates, serum, plasma, and urine. It contains Sf 21-expressed recombinant human IL-12 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate recombinant human IL-12 accurately. Results obtained using natural human IL-12 showed linear curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-12.

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.
Urine
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean25.720243619.9115223
Standard Deviation1.64.810.11.98.313.2
CV%6.22.42.39.57.25.9

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean21.222844721126246
Standard Deviation0.33.54.91.55.68.1
CV%1.41.51.17.14.43.3

Recovery

The recovery of human IL-12 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) 90 81-96
Citrate Plasma (n=5) 90 82-94
EDTA Plasma (n=5) 101 96-109
Heparin Plasma (n=5) 99 89-109
Serum (n=5) 100 89-106
Urine (n=5) 89 85-92
Linearity
To assess the linearity of the assay, five samples were spiked with high concentrations of IL-12 in various matrices and diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human IL-12 p70 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: IL-12 p70

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 p70
    • Entrez Gene IDs
      3592 (Human); 16159 (Mouse); 84405 (Rat);
    • Alternate Names
      CLMF p35; Cytotoxic lymphocyte maturation factor 35 kDa subunit; IL12 p70; IL-12 p70; IL-12 subunit p35; IL-12, subunit p35; interleukin 12, p35; interleukin 12A (natural killer cell stimulatory factor 1, cytotoxic lymphocytematuration factor 1, p35); interleukin-12 alpha chain; interleukin-12 subunit alpha; natural killer cell stimulatory factor 1, 35 kD subunit; NF cell stimulatory factor chain 1; NK cell stimulatory factor chain 1; NKSF1; p35;
    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, working standards, 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 of the Appropriate Assay Diluent
    4.   Add 50 µL of the appropriate Assay Diluent to each well.

    5. 200 µL Standard, control, or sample
    6.   Add 200 µL of Standard, control, or sample to each well. Cover with a plate sealer.
    7.   For Cell Culture Supernate, Serum & Plasma Samples:  Incubate at room temperature for 2 hours.
      For Urine Samples:  Incubate at room temperature for 1.5 hours.
    8.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.

    9. 200 µL Conjugate
    10.   Add 200 µL of Conjugate to each well, and cover with a new plate sealer.
    11.   For Cell Culture Supernate, Serum & Plasma Samples:  Incubate at room temperature for 2 hours.
      For Urine Samples:  Incubate at room temperature for 1.5 hours.
    12.   Aspirate and wash 3 times.

    13. 200 µL Substrate Solution
    14. Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.

    15. 50 µL Stop Solution
    16. 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.
    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.

    44 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. Defining the Pharmacodynamic Profile and Therapeutic Index of NHS-IL12 Immunocytokine in Dogs with Malignant Melanoma.
      Authors: Paoloni M, Mazcko C, Selting K, Lana S, Barber L, Phillips J, Skorupski K, Vail D, Wilson H, Biller B, Avery A, Kiupel M, LeBlanc A, Bernhardt A, Brunkhorst B, Tighe R, Khanna C
      PLoS ONE, 2015;10(6):e0129954.
      Species: Human
      Sample Type: Serum
    2. Endocannabinoids affect innate immunity of Muller glia during HIV-1 Tat cytotoxicity.
      Authors: Krishnan G, Chatterjee N
      Mol Cell Neurosci, 2014;59(0):10-23.
      Species: Human
      Sample Type: Cell Culture Supernates
    3. Live attenuated B. pertussis BPZE1 rescues the immune functions of Respiratory Syncytial virus infected human dendritic cells by promoting Th1/Th17 responses.
      Authors: Schiavoni I, Fedele G, Quattrini A, Bianco M, Schnoeller C, Openshaw P, Locht C, Ausiello C
      PLoS ONE, 2014;9(6):e100166.
      Species: Human
      Sample Type: Cell Culture Supernates
    4. Analysis of plasma cytokines and angiogenic factors in patients with pretreated urothelial cancer receiving Pazopanib: the role of circulating interleukin-8 to enhance the prognostic accuracy.
      Authors: Necchi A, Pennati M, Zaffaroni N, Landoni E, Giannatempo P, Raggi D, Schwartz L, Morosi C, Crippa F, Fare E, Nicolai N, Lanocita R, Sava T, Sacco C, Messina C, Ortega C, De Braud F, Salvioni R, Daidone M, Gianni A, Mariani L
      Br J Cancer, 2014;110(1):26-33.
      Species: Human
      Sample Type: Plasma
    5. Bile acids PKA-dependently induce a switch of the IL-10/IL-12 ratio and reduce proinflammatory capability of human macrophages.
      Authors: Haselow K, Bode J, Wammers M, Ehlting C, Keitel V, Kleinebrecht L, Schupp A, Haussinger D, Graf D
      J Leukoc Biol, 2013;94(6):1253-64.
      Species: Human
      Sample Type: Cell Culture Supernates
    6. Efficacy of oncolytic adenovirus expressing suicide genes and interleukin-12 in preclinical model of prostate cancer.
      Authors: Freytag, S O, Barton, K N, Zhang, Y
      Gene Ther, 2013;20(12):1131-9.
      Species: Human
      Sample Type: Cell Culture Supernates
    7. Effects of TLR agonists on maturation and function of 3-day dendritic cells from AML patients in complete remission.
      Authors: Beck B, Dorfel D, Lichtenegger FS, Geiger C, Lindner L, Merk M, Schendel DJ, Subklewe M
      J Transl Med, 2011;9(0):151.
      Species: Human
      Sample Type: Cell Culture Supernates
    8. HDAC inhibition by LBH589 affects the phenotype and function of human myeloid dendritic cells.
      Authors: Song W, Tai YT, Tian Z, Hideshima T, Chauhan D, Nanjappa P, Exley MA, Anderson KC, Munshi NC
      Leukemia, 2011;25(1):161-8.
      Species: Human
      Sample Type: Cell Culture Supernates
    9. Lack of anti-tumour reactivity despite enhanced numbers of circulating natural killer T cells in two patients with metastatic renal cell carcinoma.
      Authors: Vyth-Dreese FA, Sein J, van de Kasteele W, Dellemijn TA, van den Bogaard C, Nooijen WJ, de Gast GC, Haanen JB, Bex A
      Clin. Exp. Immunol., 2010;162(3):447-59.
      Species: Human
      Sample Type: Plasma
    10. CD14-independent responses induced by a synthetic lipid A mimetic.
      Authors: Legat A, Thomas S, Hermand P, Van Mechelen M, Goldman M, De Wit D
      Eur J Immunol, 2010;40(3):797-802.
      Species: Human
      Sample Type: Cell Culture Supernates
    11. Immunologic markers as predictors of tuberculosis-associated immune reconstitution inflammatory syndrome in HIV and tuberculosis coinfected persons in Thailand.
      Authors: Tieu HV, Ananworanich J, Avihingsanon A, Apateerapong W, Sirivichayakul S, Siangphoe U, Klongugkara S, Boonchokchai B, Hammer SM, Manosuthi W
      AIDS Res. Hum. Retroviruses, 2009;25(11):1083-9.
      Species: Human
      Sample Type: Serum
    12. Cerebrospinal fluid levels of interleukin-6 and interleukin-12 in children with meningitis.
      Authors: Hsieh CC, Lu JH, Chen SJ, Lan CC, Chow WC, Tang RB
      Cytotechnology, 2009;25(4):461-5.
      Species: Human
      Sample Type: CSF
    13. 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
    14. 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
    15. In vitro induction of a dendritic cell phenotype in primary human acute myelogenous leukemia (AML) blasts alters the chemokine release profile and increases the levels of T cell chemotactic CCL17 and CCL22.
      Authors: Olsnes AM, Ryningen A, Ersvaer E, Bruserud Ø
      J. Interferon Cytokine Res., 2008;28(5):297-310.
      Species: Human
      Sample Type: Cell Culture Supernates
    16. Translocation of bacterial DNA from Gram-positive microorganisms is associated with a species-specific inflammatory response in serum and ascitic fluid of patients with cirrhosis.
      Authors: Frances R, Gonzalez-Navajas JM, Zapater P, Munoz C, Cano R, Pascual S, Santana F, Marquez D, Perez-Mateo M, Such J
      Clin. Exp. Immunol., 2007;150(2):230-7.
      Species: Human
      Sample Type: Serum
    17. Fucoidan ingestion increases the expression of CXCR4 on human CD34+ cells.
      Authors: Irhimeh MR, Fitton JH, Lowenthal RM
      Exp. Hematol., 2007;35(6):989-94.
      Species: Human
      Sample Type: Plasma
    18. Phase 2 study of pegylated liposomal doxorubicin in combination with interleukin-12 for AIDS-related Kaposi sarcoma.
      Authors: Little RF, Aleman K, Kumar P, Wyvill KM, Pluda JM, Read-Connole E, Wang V, Pittaluga S, Catanzaro AT, Steinberg SM, Yarchoan R
      Blood, 2007;110(13):4165-71.
      Species: Human
      Sample Type: Serum
    19. 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.
    20. 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
    21. C-reactive protein and vascular cell adhesion molecule-1 as markers of severity in sickle cell disease.
      Authors: Makis AC, Hatzimichael EC, Stebbing J, Bourantas KL
      Arch. Intern. Med., 2006;166(3):366-8.
      Species: Human
      Sample Type: Plasma
    22. Celecoxib up-regulates the expression of the zeta chain of T cell receptor complex in tumor-infiltrating lymphocytes in human cervical cancer.
      Authors: Ferrandina G, Ranelletti FO, Legge F, Salutari V, Martinelli E, Fattorossi A, Lorusso D, Zannoni G, Vellone V, Paglia A, Scambia G
      Clin. Cancer Res., 2006;12(7):2055-60.
      Species: Human
      Sample Type: Plasma
    23. 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
    24. 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
    25. Chemokines indicate allergic bronchopulmonary aspergillosis in patients with cystic fibrosis.
      Authors: Hartl D, Latzin P, Zissel G, Krane M, Krauss-Etschmann S, Griese M
      Am. J. Respir. Crit. Care Med., 2006;173(12):1370-6.
      Species: Human
      Sample Type: Serum
    26. Interleukin-12 and interleukin-16 in periodontal disease.
      Authors: Tsai IS, Tsai CC, Ho YP, Ho KY, Wu YM, Hung CC
      Cytokine, 2005;31(1):34-40.
      Species: Human
      Sample Type: Gingival Crevicular Fluid (GCF)
    27. Effect of piceatannol in human monocyte-derived dendritic cells in vitro.
      Authors: Takei M, Umeyama A, Arihara S, Matsumoto H
      J Pharm Sci, 2005;94(5):974-82.
      Species: Human
      Sample Type: Cell Culture Supernates
    28. Manipulation of host cytokine network by ticks: a potential gateway for pathogen transmission.
      Authors: Hajnicka V, Vancova I, Kocakova P, Slovak M, Gasperik J, Slavikova M, Hails RS, Labuda M, Nuttall PA
      Parasitology, 2005;130(0):333-42.
      Species: Human
      Sample Type: Saliva
    29. 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
    30. Effects of antiasthmatic agents on the functions of peripheral blood monocyte-derived dendritic cells from atopic patients.
      Authors: Saeki S, Matsuse H, Kondo Y, Machida I, Kawano T, Tomari S, Obase Y, Fukushima C, Kohno S
      J. Allergy Clin. Immunol., 2004;114(3):538-44.
      Species: Human
      Sample Type: Cell Culture Supernates
    31. Reduced levels of transforming growth factor-beta1, interleukin-12 and increased migration inhibitory factor are associated with severe malaria.
      Authors: Chaiyaroj SC, Rutta AS, Muenthaisong K, Watkins P, Na Ubol M, Looareesuwan S
      Acta Trop., 2004;89(3):319-27.
      Species: Human
      Sample Type: Plasma
    32. Inflammatory cytokine concentrations are elevated in seminal plasma of men with spinal cord injuries.
      Authors: Basu S, Aballa TC, Ferrell SM, Lynne CM, Brackett NL
      J. Androl., 2004;25(2):250-4.
      Species: Human
      Sample Type: Seminal Plasma
    33. Human plasmacytoid dendritic cells activated by CpG oligodeoxynucleotides induce the generation of CD4+CD25+ regulatory T cells.
      Authors: Moseman EA, Liang X, Dawson AJ, Panoskaltsis-Mortari A, Krieg AM, Liu YJ, Blazar BR, Chen W
      J. Immunol., 2004;173(7):4433-42.
      Species: Human
      Sample Type: Cell Culture Supernates
    34. Lack of phenotypic and functional impairment in dendritic cells from chimpanzees chronically infected with hepatitis C virus.
      Authors: Larsson M, Babcock E, Grakoui A, Shoukry N, Lauer G, Rice C, Walker C, Bhardwaj N
      J. Virol., 2004;78(12):6151-61.
      Species: Primate - Pan troglodytes (Chimpanzee)
      Sample Type: Cell Culture Supernates
    35. NK cells regulate CD8+ T cell effector function in response to an intracellular pathogen.
      Authors: Vankayalapati R, Klucar P, Wizel B, Weis SE, Samten B, Safi H, Shams H, Barnes PF
      J. Immunol., 2004;172(1):130-7.
      Species: Human
      Sample Type: Cell Culture Supernates
    36. Cytokines evaluation in nasal lavage of allergic children after Bacillus clausii administration: a pilot study.
      Authors: Ciprandi G, Tosca MA, Milanese M, Caligo G, Ricca V
      Pediatr Allergy Immunol, 2004;15(2):148-51.
      Species: Human
      Sample Type: Nasal Lavage
    37. Production of interferon gamma in respiratory syncytial virus infection of humans is not associated with interleukins 12 and 18.
      Authors: Garofalo RP, Hintz KH, Hill V, Ogra PL, Welliver RC
      J. Med. Virol., 2004;73(2):289-94.
      Species: Human
      Sample Type: Tissue Secretion
    38. Local immunostimulation induced by intravesical administration of autologous interferon-gamma-activated macrophages in patients with superficial bladder cancer.
      Authors: Pages F, Lebel-Binay S, Vieillefond A, Deneux L, Cambillau M, Soubrane O, Debre B, Tardy D, Lemonne JL, Abastado JP, Fridman WH, Thiounn N
      Clin. Exp. Immunol., 2002;127(2):303-9.
      Species: Human
      Sample Type: Urine
    39. 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
    40. Intraperitoneal bispecific antibody (HEA125xOKT3) therapy inhibits malignant ascites production in advanced ovarian carcinoma.
      Authors: Marme A, Strauss G, Bastert G, Grischke EM, Moldenhauer G
      Int. J. Cancer, 2002;101(2):183-9.
      Species: Human
      Sample Type: Ascites Fluid
    41. Cytokine profile in Behcet's disease patients. Relationship with disease activity.
      Authors: Hamzaoui K, Hamzaoui A, Guemira F, Bessioud M, Hamza M, Ayed K
      Scand. J. Rheumatol., 2002;31(4):205-10.
      Species: Human
      Sample Type: Serum
    42. Interleukin-11 induces Th2 polarization of human CD4(+) T cells.
      Authors: Curti A, Ratta M, Corinti S, Girolomoni G, Ricci F, Tazzari P, Siena M, Grande A, Fogli M, Tura S, Lemoli RM
      Blood, 2001;97(9):2758-63.
      Species: Human
      Sample Type: Cell Culture Supernates
    43. Mature dendritic cells infiltrate the T cell-rich region of oral mucosa in chronic periodontitis: in situ, in vivo, and in vitro studies.
      Authors: Jotwani R, Palucka AK, Al-Quotub M, Nouri-Shirazi M, Kim J, Bell D, Banchereau J, Cutler CW
      J. Immunol., 2001;167(8):4693-700.
      Species: Human
      Sample Type: Gingival Crevicular Fluid (GCF)
    44. Natural interferon alpha/beta-producing cells link innate and adaptive immunity.
      Authors: Kadowaki N, Antonenko S, Lau JY, Liu YJ
      J. Exp. Med., 2000;192(2):219-26.
      Species: Human
      Sample Type: Cell Culture Supernates
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