Mouse IL-12/IL-23 p40 Allele-specific DuoSet ELISA

Catalog # Availability Size / Price Qty
DY499
DY499-05
Ancillary Products Available
IL-12/IL-23 p40 ELISA Kit ELISA
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Citations (39)
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Mouse IL-12/IL-23 p40 Allele-specific 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 mouse IL-12 p40 (allele-specific). 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

Mouse IL-12 / IL-23 p40 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/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:
CLMF p40; CLMF; CLMF2; Cytotoxic lymphocyte maturation factor 40 kDa subunit; IL12 p40; IL-12 p40; IL-12 subunit p40; IL12B; IL-12B; IL-12BNK cell stimulatory factor chain 2; interleukin 12, p40; interleukin 12B (natural killer cell stimulatory factor 2, cytotoxic lymphocytematuration factor 2, p40); interleukin-12 beta chain; interleukin-12 subunit beta; natural killer cell stimulatory factor, 40 kD subunit; NKSF; NKSF2; NKSF2IL12, subunit p40

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.

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 Mouse IL-12/IL-23 p40 Allele-specific 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.

39 Citations: Showing 1 - 10
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  1. IL-23R Signaling Plays No Role in Myocardial Infarction
    Authors: E Engelowski, NF Modares, S Gorressen, P Bouvain, D Semmler, C Alter, Z Ding, U Flögel, J Schrader, H Xu, PA Lang, J Fischer, DM Floss, J Scheller
    Sci Rep, 2018;8(1):17078.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  2. Interferon induced protein 35 exacerbates H5N1 influenza disease through the expression of IL-12p40 homodimer
    Authors: AP Gounder, CC Yokoyama, NN Jarjour, TL Bricker, BT Edelson, ACM Boon
    PLoS Pathog., 2018;14(4):e1007001.
    Species: Mouse
    Sample Types: Serum
  3. Intravesicular Acidification Regulates Lipopolysaccharide Inflammation and Tolerance through TLR4 Trafficking
    Authors: M Murase, T Kawasaki, R Hakozaki, T Sueyoshi, DDP Putri, Y Kitai, S Sato, M Ikawa, T Kawai
    J. Immunol., 2018;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  4. IRF5 distinguishes severe asthma in humans and drives Th1 phenotype and airway hyperreactivity in mice
    Authors: TB Oriss, M Raundhal, C Morse, RE Huff, S Das, R Hannum, MC Gauthier, KL Scholl, K Chakrabort, SM Nouraie, SE Wenzel, P Ray, A Ray
    JCI Insight, 2017;2(10):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  5. Necroptosis promotes autophagy-dependent upregulation of DAMP and results in immunosurveillance
    Authors: SY Lin, SY Hsieh, YT Fan, WC Wei, PW Hsiao, DH Tsai, TS Wu, NS Yang
    Autophagy, 2017;0(0):0.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  6. Surface layer proteins from virulent Clostridium difficile ribotypes exhibit signatures of positive selection with consequences for innate immune response
    Authors: M Lynch, TA Walsh, I Marszalows, AE Webb, M MacAogain, TR Rogers, H Windle, D Kelleher, MJ O'Connell, CE Loscher
    BMC Evol. Biol, 2017;17(1):90.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  7. MyD88 adapter-like (Mal)/TIRAP is required for cytokine production by splenic Ly6CloTLR2hi but not by Ly6ChiTLR2hi monocytes during Trypanosoma cruzi Infection
    J Biol Chem, 2016;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  8. Lectins from Synadenium carinatum (ScLL) and Artocarpus heterophyllus (ArtinM) Are Able to Induce Beneficial Immunomodulatory Effects in a Murine Model for Treatment of Toxoplasma gondii Infection
    Authors: , EL Ramos, SS Santana, MV Silva, FM Santiago, TW Mineo, JR Mineo
    Front Cell Infect Microbiol, 2016;6(0):164.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  9. Alternatively Activated Mononuclear Phagocytes from the Skin Site of Infection and the Impact of IL-4R? Signalling on CD4+T Cell Survival in Draining Lymph Nodes after Repeated Exposure to Schistosoma mansoni Cercariae
    PLoS Negl Trop Dis, 2016;10(8):e0004911.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  10. Overexpression of IL-15 promotes tumor destruction via NK1.1+ cells in a spontaneous breast cancer model.
    Authors: Gillgrass A, Chew M, Krneta T, Ashkar A
    BMC Cancer, 2015;15(0):293.
    Species: Mouse
    Sample Types: Tissue Homogenates
  11. Non-canonical interleukin 23 receptor complex assembly: p40 protein recruits interleukin 12 receptor beta1 via site II and induces p19/interleukin 23 receptor interaction via site III.
    Authors: Schroder J, Moll J, Baran P, Grotzinger J, Scheller J, Floss D
    J Biol Chem, 2015;290(1):359-70.
  12. MyD88 signalling in myeloid cells is sufficient to prevent chronic mycobacterial infection.
    Authors: Berod L, Stuve P, Swallow M, Arnold-Schrauf C, Kruse F, Gentilini M, Freitag J, Holzmann B, Sparwasser T
    Eur J Immunol, 2014;44(5):1399-409.
    Species: Mouse
    Sample Types: Serum
  13. Development and characterization of a long-term murine model of Streptococcus pneumoniae infection of the lower airways.
    Authors: Haste L, Hulland K, Bolton S, Yesilkaya H, McKechnie K, Andrew P
    Infect Immun, 2014;82(8):3289-98.
    Species: Mouse
    Sample Types: BALF
  14. Intraperitoneal prophylaxis with CpG oligodeoxynucleotides protects neutropenic mice against intracerebral Escherichia coli K1 infection.
    Authors: Ribes S, Meister T, Ott M, Redlich S, Janova H, Hanisch U, Nessler S, Nau R
    J Neuroinflammation, 2014;11(0):14.
    Species: Mouse
    Sample Types: Tissue Homogenates
  15. TLR9 and MyD88 are crucial for the development of protective immunity to malaria.
    Authors: Gowda NM, Wu X, Gowda DC
    J. Immunol., 2012;188(10):5073-85.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  16. Macrophage Dectin-1 Expression Is Controlled by Leukotriene B4 via a GM-CSF/PU.1 Axis.
    Authors: Serezani CH, Kane S, Collins L
    J. Immunol., 2012;189(2):906-15.
    Species: Mouse
    Sample Types: BALF
  17. CD4 T cells promote rather than control tuberculosis in the absence of PD-1-mediated inhibition.
    Authors: Barber DL, Mayer-Barber KD, Feng CG
    J. Immunol., 2011;186(3):1598-607.
    Species: Mouse
    Sample Types: BALF
  18. A critical role for C5L2 in the pathogenesis of experimental allergic asthma.
    Authors: Zhang X, Schmudde I, Laumonnier Y, Pandey MK, Clark JR, Konig P, Gerard NP, Gerard C, Wills-Karp M, Kohl J
    J. Immunol., 2010;185(11):6741-52.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  19. Chorionic gonadotropin alleviates thioglycollate-induced peritonitis by affecting macrophage function.
    Authors: Wan H, Coppens JM, van Helden-Meeuwsen CG, Leenen PJ, Van Rooijen N, Khan NA, Kiekens RC, Benner R, Versnel MA
    J. Leukoc. Biol., 2009;86(2):361-70.
    Species: Mouse
    Sample Types: Peritoneal Lavage
  20. A conjugated linoleic acid-enriched beef diet attenuates lipopolysaccharide-induced inflammation in mice in part through PPARgamma-mediated suppression of toll-like receptor 4.
    Authors: Reynolds CM, Draper E, Keogh B, Rahman A, Moloney AP, Mills KH, Loscher CE, Roche HM
    J. Nutr., 2009;139(12):2351-7.
    Species: Mouse
    Sample Types: Serum
  21. Administration of a synthetic TLR4 agonist protects mice from pneumonic tularemia.
    Authors: Lembo A, Pelletier M, Iyer R, Timko M, Dudda JC, West TE, Wilson CB, Hajjar AM, Skerrett SJ
    J. Immunol., 2008;180(11):7574-81.
    Species: Mouse
    Sample Types: Tissue Homogenates
  22. Decreased pathology and prolonged survival of human DC-SIGN transgenic mice during mycobacterial infection.
    Authors: Schaefer M, Reiling N, Fessler C, Stephani J, Taniuchi I, Hatam F, Yildirim AO, Fehrenbach H, Walter K, Ruland J, Wagner H, Ehlers S, Sparwasser T
    J. Immunol., 2008;180(10):6836-45.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  23. Altered inflammatory responses in TLR5-deficient mice infected with Legionella pneumophila.
    Authors: Hawn TR, Berrington WR, Smith IA, Uematsu S, Akira S, Aderem A, Smith KD, Skerrett SJ
    J. Immunol., 2007;179(10):6981-7.
    Species: Mouse
    Sample Types: Tissue Homogenates
  24. Differential involvement of TLR2 and TLR4 in host survival during pulmonary infection with Chlamydia pneumoniae.
    Authors: Rodriguez N, Wantia N, Fend F, Durr S, Wagner H, Miethke T
    Eur. J. Immunol., 2006;36(5):1145-55.
    Species: Mouse
    Sample Types: Tissue Homogenates
  25. TLR4 mediates vaccine-induced protective cellular immunity to Bordetella pertussis: role of IL-17-producing T cells.
    Authors: Higgins SC, Jarnicki AG, Lavelle EC, Mills KH
    J. Immunol., 2006;177(11):7980-9.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  26. The acylation state of mycobacterial lipomannans modulates innate immunity response through toll-like receptor 2.
    Authors: Gilleron M, Nigou J, Nicolle D, Quesniaux V, Puzo G
    Chem. Biol., 2006;13(1):39-47.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  27. MyD88-dependent and -independent murine cytomegalovirus sensing for IFN-alpha release and initiation of immune responses in vivo.
    Authors: Delale T, Paquin A, Asselin-Paturel C, Dalod M, Brizard G, Bates EE, Kastner P, Chan S, Akira S, Vicari A, Biron CA, Trinchieri G, Briere F
    J. Immunol., 2005;175(10):6723-32.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  28. Both hemopoietic and resident cells are required for MyD88-dependent pulmonary inflammatory response to inhaled endotoxin.
    Authors: Noulin N, Quesniaux VF, Schnyder-Candrian S, Schnyder B, Maillet I, Robert T, Vargaftig BB, Ryffel B, Couillin I
    J. Immunol., 2005;175(10):6861-9.
    Species: Mouse
    Sample Types: BALF
  29. Simvastatin augments lipopolysaccharide-induced proinflammatory responses in macrophages by differential regulation of the c-Fos and c-Jun transcription factors.
    Authors: Matsumoto M, Einhaus D, Gold ES, Aderem A
    J. Immunol., 2004;172(12):7377-84.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  30. Stimulation of toll-like receptor 2 by Coxiella burnetii is required for macrophage production of pro-inflammatory cytokines and resistance to infection.
    Authors: Zamboni DS, Campos MA, Torrecilhas AC, Kiss K, Samuel JE, Golenbock DT, Lauw FN, Roy CR, Almeida IC, Gazzinelli RT
    J. Biol. Chem., 2004;279(52):54405-15.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  31. Toll-like receptor 2 is required for optimal control of Listeria monocytogenes infection.
    Authors: Torres D, Barrier M, Bihl F, Quesniaux VJ, Maillet I, Akira S, Ryffel B, Erard F
    Infect. Immun., 2004;72(4):2131-9.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  32. Fatal Mycobacterium tuberculosis infection despite adaptive immune response in the absence of MyD88.
    Authors: Fremond CM, Yeremeev V, Nicolle DM, Jacobs M, Quesniaux VF, Ryffel B
    J. Clin. Invest., 2004;114(12):1790-9.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  33. Long-term control of Mycobacterium bovis BCG infection in the absence of Toll-like receptors (TLRs): investigation of TLR2-, TLR6-, or TLR2-TLR4-deficient mice.
    Authors: Nicolle D, Fremond C, Pichon X, Bouchot A, Maillet I, Ryffel B, Quesniaux VJ
    Infect. Immun., 2004;72(12):6994-7004.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  34. Severe pancreatitis with exocrine destruction and increased islet neogenesis in mice with suppressor of cytokine signaling-1 deficiency.
    Authors: Chong MM, Thomas HE, Kay TW
    Am. J. Pathol., 2004;165(3):913-21.
    Species: Mouse
    Sample Types: Serum
  35. Cytokine production in association with phagocytosis of apoptotic cells by immature dendritic cells.
    Authors: Takahashi M, Kobayashi Y
    Cell. Immunol., 2003;226(2):105-15.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  36. Acylation state of the phosphatidylinositol hexamannosides from Mycobacterium bovis bacillus Calmette Guerin and mycobacterium tuberculosis H37Rv and its implication in Toll-like receptor response.
    Authors: Gilleron M, Quesniaux VF, Puzo G
    J. Biol. Chem., 2003;278(32):29880-9.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  37. A role for tumor necrosis factor alpha in death of dopaminergic neurons following neural transplantation.
    Authors: Clarke DJ, Branton RL
    Exp. Neurol., 2002;176(1):154-62.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  38. CpG-DNA aided cross-presentation of soluble antigens by dendritic cells.
    Authors: O'Keeffe M
    Eur. J. Immunol., 2002;32(8):2356-64.
    Species: Mouse
    Sample Types: Serum
  39. Reversal of tumor-induced dendritic cell paralysis by CpG immunostimulatory oligonucleotide and anti-interleukin 10 receptor antibody.
    Authors: O'Garra A
    J. Exp. Med., 2002;196(4):541-9.
    Species: Mouse
    Sample Types: Cell Culture Supernates

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