Human IL-12 p70 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY1270
DY1270-05
Ancillary Products Available
IL-12 p70 ELISA Kit ELISA
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Product Details
Procedure
Citations (36)
FAQs
Supplemental Products
Reviews (6)

Human IL-12 p70 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human IL-12 p70. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

Normal Goat Serum: (Catalog # DY005)

 

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Normal Goat Serum: (Catalog # DY005)

 

Data Example

Human IL-12 p70 ELISA Standard Curve

Product Datasheets

Preparation and Storage

Stability & 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

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent with NGS, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human IL-12 p70 DuoSet ELISA

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.

36 Citations: Showing 1 - 10
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  1. Plasmodium falciparum Treated with Artemisinin-based Combined Therapy Exhibits Enhanced Mutation, Heightened Cortisol and TNF-? Induction
    Authors: AO Idowu, S Bhattachar, S Gradus, W Oyibo, Z George, C Black, J Igietseme, AA Azenabor
    Int J Med Sci, 2018;15(13):1449-1457.
    Species: Human
    Sample Types: Plasma
  2. Interferon priming is essential for human CD34+ cell-derived plasmacytoid dendritic cell maturation and function
    Authors: A Laustsen, RO Bak, C Krapp, L Kjær, JH Egedahl, CC Petersen, S Pillai, HQ Tang, N Uldbjerg, M Porteus, NR Roan, M Nyegaard, PW Denton, MR Jakobsen
    Nat Commun, 2018;9(1):3525.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. A novel susceptibility locus in the IL12B region is associated with the pathophysiology of Takayasu arteritis through IL-12p40 and IL-12p70 production
    Authors: T Nakajima, H Yoshifuji, M Shimizu, K Kitagori, K Murakami, R Nakashima, Y Imura, M Tanaka, K Ohmura, F Matsuda, C Terao, T Mimori
    Arthritis Res. Ther., 2017;19(1):197.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Dexamethasone induced inhibition of Dectin-1 activation of antigen presenting cells is mediated via STAT-3 and NF-?B signaling pathways.
    Authors: Philipp Kotthoff, Annkristin Heine, Stefanie Andrea Erika Held, Peter Brossart
    Scientific Reports, 2017;0(0):2045-2322.
    Species: Human
    Sample Types: Cell Culture Supernate
  5. Th1/Th17-Related Cytokines and Chemokines and Their Implications in the Pathogenesis of Pemphigus Vulgaris
    Authors: RP Timoteo, MV da Silva, CB Miguel, DA Silva, JD Catarino, V Rodrigues, H Sales-Camp, CJ Freire Oli
    Mediators Inflamm., 2017;2017(0):7151285.
    Species: Human
    Sample Types: Serum
  6. Minimal hepatic encephalopathy is associated with expansion and activation of CD(4+)CD28(-), Th22 and Tfh and B lymphocytes
    Authors: A Mangas-Los, R García-Gar, A Urios, D Escudero-G, J Tosca, R Giner-Durá, MA Serra, C Montoliu, V Felipo
    Sci Rep, 2017;7(1):6683.
    Species: Human
    Sample Types: Serum
  7. The effects of exogenous lipid on THP-1 cells: an in vitro model of airway aspiration?
    Authors: YA Hayman, LR Sadofsky, JD Williamson, SP Hart, AH Morice
    ERJ Open Res, 2017;3(1):.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Acanthamoeba castellanii (Genotype T4) Stimulates the Production of Interleukin-10 as well as Pro-inflammatory Cytokines in THP-1 Cells, Human Peripheral Blood Mononuclear Cells and Human Monocyte-Derived Macrophages
    Infect Immun, 2016;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Transcriptional and functional characterization of CD137L-dendritic cells identifies a novel dendritic cell phenotype
    Sci Rep, 2016;6(0):29712.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Exposure to diesel exhaust particle extracts (DEPe) impairs some polarization markers and functions of human macrophages through activation of AhR and Nrf2.
    Authors: Jaguin M, Fardel O, Lecureur V
    PLoS ONE, 2015;10(2):e0116560.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Paracoccidioides brasiliensis interferes on dendritic cells maturation by inhibiting PGE2 production.
    Authors: Fernandes R, Bachiega T, Rodrigues D, Golim M, Dias-Melicio L, Balderramas H, Kaneno R, Soares A
    PLoS ONE, 2015;10(3):e0120948.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. The effects of cytokines on spontaneous hepatitis B surface antigen seroconversion in chronic hepatitis B virus infection.
    Authors: Wu J, Hsu H, Chiu Y, Chen H, Ni Y, Chang M
    J Immunol, 2015;194(2):690-6.
    Species: Human
    Sample Types: Serum
  13. Priming of Human Resting NK Cells by Autologous M1 Macrophages via the Engagement of IL-1beta, IFN-beta, and IL-15 Pathways.
    Authors: Mattiola I, Pesant M, Tentorio P, Molgora M, Marcenaro E, Lugli E, Locati M, Mavilio D
    J Immunol, 2015;195(6):2818-28.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. Macrophages and dendritic cells as actors in the immune reaction of classical Hodgkin lymphoma.
    Authors: Tudor C, Bruns H, Daniel C, Distel L, Hartmann A, Gerbitz A, Buettner M
    PLoS ONE, 2015;9(12):e114345.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Anti-inflammatory effects of green soybean extract irradiated with visible light.
    Authors: Tanaka K, Ohgo Y, Katayanagi Y, Yasui K, Hiramoto S, Ikemoto H, Nakata Y, Miyoshi N, Isemura M, Ohashi N, Imai S
    Sci Rep, 2014;4(0):4732.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. Two wheat decapeptides prevent gliadin-dependent maturation of human dendritic cells.
    Authors: Giordani L, Del Pinto T, Vincentini O, Felli C, Silano M, Viora M
    Exp Cell Res, 2014;321(2):248-54.
    Species: Human
    Sample Types: Cell Culture Supernates
  17. Tumour microenvironment of both early- and late-stage colorectal cancer is equally immunosuppressive.
    Authors: O'Toole A, Michielsen A, Nolan B, Tosetto M, Sheahan K, Mulcahy H, Winter D, Hyland J, O'Connell P, Fennelly D, O'Donoghue D, O'Sullivan J, Doherty G, Ryan E
    Br J Cancer, 2014;111(5):927-32.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. Effect of CIK on multidrug-resistance reversal and increasing the sensitivity of ADR in K562/ADR cells.
    Authors: Wang L, Deng Q, Wang J, Bai X, Xiao X, Lv H, Zhao M, Liu P
    Oncol Lett, 2014;8(4):1778-1782.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. Expression and functional characterization of NOD2 in decidual stromal cells isolated during the first trimester of pregnancy.
    Authors: Zhang Y, Chen H, Sun C, Wang H, Liu M, Li Y, Nie X, Du M, Li D, Zhang J
    PLoS ONE, 2014;9(6):e99612.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. CD137 ligand signalling induces differentiation of primary acute myeloid leukaemia cells.
    Authors: Cheng K, Wong S, Linn Y, Ho L, Chng W, Schwarz H
    Br J Haematol, 2014;165(1):134-44.
    Species: Human
    Sample Types: Cell Culture Supernates
  21. Selective activation of human dendritic cells by OM-85 through a NF-kB and MAPK dependent pathway.
    Authors: Parola C, Salogni L, Vaira X, Scutera S, Somma P, Salvi V, Musso T, Tabbia G, Bardessono M, Pasquali C, Mantovani A, Sozzani S, Bosisio D
    PLoS ONE, 2013;8(12):e82867.
    Species: Human
    Sample Types: Cell Culture Supernates
  22. Mechanisms inducing low bone density in duchenne muscular dystrophy in mice and humans.
    Authors: Rufo A, Del Fattore A, Capulli M, Carvello F, De Pasquale L, Ferrari S, Pierroz D, Morandi L, De Simone M, Rucci N, Bertini E, Bianchi ML, De Benedetti F, Teti A
    J. Bone Miner. Res., 2011;26(8):1891-903.
    Species: Human
    Sample Types: Serum
  23. Human uterine epithelial cell secretions regulate dendritic cell differentiation and responses to TLR ligands.
    Authors: Ochiel DO, Ghosh M, Fahey JV
    J. Leukoc. Biol., 2010;88(3):435-44.
    Species: Human
    Sample Types: Cell Culture Supernates
  24. Expression of soluble vascular endothelial growth factor receptor-1 in human monocyte-derived mature dendritic cells contributes to their antiangiogenic property.
    Authors: Kishuku M, Nishioka Y, Abe S, Kishi J, Ogino H, Aono Y, Azuma M, Kinoshita K, Batmunkh R, Rentsenhand B, Makino H, Ranjan P, Minakuchi K, Sone S
    J. Immunol., 2009;183(12):8176-85.
    Species: Human
    Sample Types: Cell Culture Supernates
  25. Interleukin-17 and interferon-gamma are produced concomitantly by human coronary artery-infiltrating T cells and act synergistically on vascular smooth muscle cells.
    Authors: Eid RE, Rao DA, Zhou J, Lo SF, Ranjbaran H, Gallo A, Sokol SI, Pfau S, Pober JS, Tellides G
    Circulation, 2009;119(10):1424-32.
    Species: Human
    Sample Types: Plasma
  26. 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 Types: Serum
  27. Lactoferrin, a major defense protein of innate immunity, is a novel maturation factor for human dendritic cells.
    Authors: Spadaro M, Caorsi C, Ceruti P, Varadhachary A, Forni G, Pericle F, Giovarelli M
    FASEB J., 2008;22(8):2747-57.
    Species: Human
    Sample Types: Cell Culture Supernates
  28. Tumor-associated leukemia inhibitory factor and IL-6 skew monocyte differentiation into tumor-associated macrophage-like cells.
    Authors: Duluc D, Delneste Y, Tan F, Moles MP, Grimaud L, Lenoir J, Preisser L, Anegon I, Catala L, Ifrah N, Descamps P, Gamelin E, Gascan H, Hebbar M, Jeannin P
    Blood, 2007;110(13):4319-30.
    Species: Human
    Sample Types: Cell Culture Supernates
  29. The differential effect of Eriobotrya japonica hydrophilic leaf extract on cytokines production and modulation.
    Authors: Matalka KZ, Ali D, Khawad AE, Qa'dan F
    Cytokine, 2007;40(3):235-40.
    Species: Human
    Sample Types: Cell Culture Supernates
  30. Human epithelial cells establish direct antifungal defense through TLR4-mediated signaling.
    Authors: Weindl G, Naglik JR, Kaesler S, Biedermann T, Hube B, Korting HC, Schaller M
    J. Clin. Invest., 2007;117(12):3664-3672.
    Species: Human
    Sample Types: Cell Culture Supernates
  31. The direct effects of Toll-like receptor ligands on human NK cell cytokine production and cytotoxicity.
    Authors: Lauzon NM, Mian F, MacKenzie R, Ashkar AA
    Cell. Immunol., 2006;241(2):102-12.
    Species: Human
    Sample Types: Cell Culture Supernates
  32. A cyanobacterial LPS antagonist prevents endotoxin shock and blocks sustained TLR4 stimulation required for cytokine expression.
    Authors: Macagno A, Molteni M, Rinaldi A
    J. Exp. Med., 2006;203(6):1481-92.
    Species: Human
    Sample Types: Cell Culture Supernates
  33. Synergistic stimulation of human monocytes and dendritic cells by Toll-like receptor 4 and NOD1- and NOD2-activating agonists.
    Authors: Fritz JH, Girardin SE, Fitting C, Werts C, Mengin-Lecreulx D, Caroff M, Cavaillon JM, Philpott DJ, Adib-Conquy M
    Eur. J. Immunol., 2005;35(8):2459-70.
    Species: Human
    Sample Types: Cell Culture Supernates
  34. The Toll-like receptor-2/6 agonist macrophage-activating lipopeptide-2 cooperates with IFN-gamma to reverse the Th2 skew in an in vitro allergy model.
    Authors: Weigt H, Muhlradt PF, Larbig M, Krug N, Braun A
    J. Immunol., 2004;172(10):6080-6.
    Species: Human
    Sample Types: Cell Culture Supernates
  35. Mouse strain differences in plasmacytoid dendritic cell frequency and function revealed by a novel monoclonal antibody.
    Authors: Asselin-Paturel C, Brizard G, Pin JJ, Briere F, Trinchieri G
    J. Immunol., 2003;171(12):6466-77.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  36. Pleural Effusions from Patients with Mesothelioma Induce Recruitment of Monocytes and Their Differentiation into M2 Macrophages.
    Authors: Chene A, d'Almeida S, Blondy T, Tabiasco J, Deshayes S, Fonteneau J, Cellerin L, Delneste Y, Gregoire M, Blanquart C
    J Thorac Oncol, 0;11(10):1765-73.
    Species: Human
    Sample Types: Cell Culture Supernates

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Reviews for Human IL-12 p70 DuoSet ELISA

Average Rating: 4.3 (Based on 6 Reviews)

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Human IL-12 p70 DuoSet ELISA
By Anonymous on 02/22/2018
Sample Tested: THP-1 human acute monocytic leukemia cell line

Human IL-12 p70 DuoSet ELISA
By Anonymous on 01/18/2018
Sample Tested: EDTA Plasma

Human IL-12 p70 DuoSet ELISA
By Anonymous on 11/21/2017
Sample Tested: Serum

We used human serum at a dilution of 1:1. The test was good and worked well, but IL-12p70 was detectable in only 15% of our patients, in our conditions.


Human IL-12 p70 DuoSet ELISA
By Anonymous on 06/02/2016
Sample Tested: Human cells

High Quality Elisa we use for our release testing


Human IL-12 p70 DuoSet ELISA
By Anonymous on 04/22/2016
Sample Tested: Serum and Plasma

Human serum and plasma samples are not possible to run with this kit. All samples are lower than the lowest standard.


Human IL-12 p70 DuoSet ELISA, 15 Plate
By Anonymous on 10/26/2015
Sample Tested: Dendritic cells Species: Human

Standard curve is shown.
Buffer: kit dilution buffer
Dilution: 1/100