Mouse IL-12 p40 Allele-specific Quantikine ELISA Kt

Catalog # Availability Size / Price Qty
PM1240
SM1240
M1240
IL-12/IL-23 p40 ELISA Kit ELISA
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Product Details
Procedure
Citations (28)
FAQs
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Mouse IL-12 p40 Allele-specific Quantikine ELISA Kt Summary

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.

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

Cell Culture Supernates, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 42.2 161 588 41.9 151 581
Standard Deviation 0.81 1.7 9.6 4.1 10.3 24
CV% 1.9 1.1 1.6 9.8 6.8 4.1

Recovery

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

Linearity

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/IL-23 p40 Allele-specific ELISA Linearity

Data Examples

Mouse IL-12/IL-23 p40 Allele-specific ELISA Standard Curve

Product Datasheets

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:
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
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

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.

Citations for Mouse IL-12 p40 Allele-specific Quantikine ELISA Kt

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.

28 Citations: Showing 1 - 10
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  1. The Orphan Nuclear Receptor NR4A3 Is Involved in the Function of Dendritic Cells
    Authors: M Nagaoka, T Yashiro, Y Uchida, T Ando, M Hara, H Arai, H Ogawa, K Okumura, K Kasakura, C Nishiyama
    J. Immunol., 2017;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  2. 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 Types: Cell Culture Supernates
  3. 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 Types: Cell Culture Supernates
  4. 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 Types: Cell Culture 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 Types: Serum
  6. 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 Types: Cell Culture Supernates
  7. 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 Types: BALF
  8. 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 Types: Serum
  9. 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 Types: Serum
  10. 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 Types: Serum
  11. 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 Types: Cell Culture Supernates
  12. 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 Types: Serum
  13. 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 Types: Cell Culture Supernates
  14. 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 Types: Cell Culture Supernates
  15. 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 Types: Cell Culture Supernates
  16. 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 Types: 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: Mouse
    Sample Types: Cell Culture Supernates
  18. 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 Types: Cell Culture Supernates
  19. 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 Types: Cell Culture Supernates
  20. 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 Types: Cell Culture Supernates
  21. 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 Types: Peritoneal Fluid
  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 Types: Peritoneal Fluid
  23. 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 Types: Tissue Secretion
  24. 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 Types: BALF
  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 Types: Serum
  26. 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 Types: Tissue Homogenates
  27. 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 Types: Cell Culture Supernates
  28. 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 Types: Cell Culture Supernates

FAQs

  1. What is the difference between the Mouse IL-12 p40 Allele-specific (M1240) and Non Allele-specific(MP400) Quantikine ELISA kits?

    • Polymorphisms exist in the mouse IL-12/IL-23 p40 sequence. The monoclonal capture antibody used in M1240  is allele-specific. It recognizes the IL-12/IL-23 p40 variant with methionine at residue 169 (Accession # P43432) in mouse strains A.SW, BALB/c, C3H, DBA/1, C57BL/6, NOR, and 129/J.  The MP400 kit recognizes variant and non-variant mouse IL-12/IL-23 p40.

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