Mouse IL-12 p40 NonAllele-specific Quantikine ELISA

Catalog # Availability Size / Price Qty
MP400
SMP400
PMP400
IL-12/IL-23 p40 ELISA Kit ELISA
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Product Details
Procedure
Citations (16)
FAQs
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Mouse IL-12 p40 NonAllele-specific Quantikine ELISA 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 (25 uL), EDTA Plasma (25 uL), Heparin Plasma (25 uL)
Sensitivity
2.7 pg/mL
Assay Range
11.7 - 750 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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, mouse serum, and plasma. It contains recombinant mouse IL-12/23 p40 homodimer and antibodies raised against the p40 subunit of recombinant mouse IL-12 p70. 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, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 26 21 27
Mean 33.3 56.4 397 36.5 62.7 405
Standard Deviation 2.3 3 17.2 3.8 6.5 33.7
CV% 6.9 5.3 4.3 10.4 10.4 8.3

Recovery

The recovery of mouse IL-12/IL-23 p40 spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=4) 104 98-108
EDTA Plasma (n=4) 99 81-112
Heparin Plasma (n=4) 95 80-109
Serum (n=4) 105 86-117

Linearity

To assess the linearity of the assay, four 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 Non Allele-specific ELISA Linearity

Data Examples

Mouse IL-12/IL-23 p40 Non 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 NonAllele-specific Quantikine 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.

16 Citations: Showing 1 - 10
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  1. Modelling of mouse experimental colitis by global property screens: a holistic approach to assess drug effects in inflammatory bowel disease.
    Authors: Gottfries J, Melgar S, Michaelsson E
    PLoS ONE, 2012;7(1):e30005.
    Species: Mouse
    Sample Types: Tissue Homogenates
  2. Cutting edge: Divergent cell-specific functions of MyD88 for inflammatory responses and organ injury in septic peritonitis.
    Authors: Gais P, Reim D, Jusek G, Rossmann-Bloeck T, Weighardt H, Pfeffer K, Altmayr F, Janssen KP, Holzmann B
    J. Immunol., 2012;188(12):5833-7.
    Species: Mouse
    Sample Types: Plasma
  3. Human mesenchymal stem/stromal cells cultured as spheroids are self-activated to produce prostaglandin E2 that directs stimulated macrophages into an anti-inflammatory phenotype.
    Authors: Ylostalo J, Bartosh T, Coble K, Prockop D
    Stem Cells, 2012;30(10):2283-96.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  4. Serotonin activates dendritic cell function in the context of gut inflammation.
    Authors: Li N, Ghia JE, Wang H, McClemens J, Cote F, Suehiro Y, Mallet J, Khan WI
    Am. J. Pathol., 2011;178(2):662-71.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  5. ChemR23 dampens lung inflammation and enhances anti-viral immunity in a mouse model of acute viral pneumonia.
    Authors: Bondue B, Vosters O, de Nadai P, Glineur S, De Henau O, Luangsay S, Van Gool F, Communi D, De Vuyst P, Desmecht D, Parmentier M
    PLoS Pathog., 2011;7(11):e1002358.
    Species: Mouse
    Sample Types: Tissue Homogenates
  6. p47phox deficiency induces macrophage dysfunction resulting in progressive crystalline macrophage pneumonia.
    Authors: Liu Q, Cheng LI, Yi L, Zhu N, Wood A, Changpriroa CM, Ward JM, Jackson SH
    Am. J. Pathol., 2009;174(1):153-63.
    Species: Mouse
    Sample Types: BALF
  7. Unc93B1 biases Toll-like receptor responses to nucleic acid in dendritic cells toward DNA- but against RNA-sensing.
    Authors: Fukui R, Saitoh S, Matsumoto F, Kozuka-Hata H, Oyama M, Tabeta K, Beutler B, Miyake K
    J. Exp. Med., 2009;206(6):1339-50.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  8. Pulmonary stromal cells induce the generation of regulatory DC attenuating T-cell-mediated lung inflammation.
    Authors: Li Q, Guo Z, Xu X, Xia S, Cao X
    Eur. J. Immunol., 2008;38(10):2751-61.
    Species: Mouse
    Sample Types: BALF
  9. Cytokine production by M-CSF- and GM-CSF-induced mouse bone marrow-derived macrophages upon coculturing with late apoptotic cells.
    Authors: Yamazaki T, Nagata K, Kobayashi Y
    Cell. Immunol., 2008;251(2):124-30.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  10. Cigarette smoke selectively enhances viral PAMP- and virus-induced pulmonary innate immune and remodeling responses in mice.
    Authors: Kang MJ, Lee CG, Lee JY, Dela Cruz CS, Chen ZJ, Enelow R, Elias JA
    J. Clin. Invest., 2008;118(8):2771-84.
    Species: Mouse
    Sample Types: BALF
  11. NOD2-deficient mice have impaired resistance to Mycobacterium tuberculosis infection through defective innate and adaptive immunity.
    Authors: Divangahi M, Mostowy S, Coulombe F, Kozak R, Guillot L, Veyrier F, Kobayashi KS, Flavell RA, Gros P, Behr MA
    J. Immunol., 2008;181(10):7157-65.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  12. Toll-like receptor (TLR)2 and TLR3 synergy and cross-inhibition in murine myeloid dendritic cells.
    Authors: Vanhoutte F, Paget C, Breuilh L, Fontaine J, Vendeville C, Goriely S, Ryffel B, Faveeuw C, Trottein F
    Immunol. Lett., 2007;116(1):86-94.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  13. The transcriptional response to lipopolysaccharide reveals a role for interferon-gamma in lung neutrophil recruitment.
    Authors: Burch LH, Yang IV, Whitehead GS, Chao FG, Berman KG, Schwartz DA
    Am. J. Physiol. Lung Cell Mol. Physiol., 2006;291(4):L677-82.
    Species: Mouse
    Sample Types: BALF
  14. A novel siRNA-lipoplex technology for RNA interference in the mouse vascular endothelium.
    Authors: Santel A, Aleku M, Keil O, Endruschat J, Esche V, Fisch G, Dames S, Loffler K, Fechtner M, Arnold W, Giese K, Klippel A, Kaufmann J
    Gene Ther., 2006;13(16):1222-34.
    Species: Mouse
    Sample Types: Serum
  15. Oral administration of an immunostimulatory DNA sequence from Bifidobacterium longum improves Th1/Th2 balance in a murine model.
    Authors: Takahashi N, Kitazawa H, Iwabuchi N, Xiao JZ, Miyaji K, Iwatsuki K, Saito T
    Biosci. Biotechnol. Biochem., 2006;70(8):2013-7.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  16. Dendritic cells phagocytose and are activated by Treponema pallidum.
    Authors: Bouis DA, Popova TG, Takashima A, Norgard MV
    Infect. Immun., 2001;69(1):518-28.
    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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