Porcine IL-12/IL-23 p40 Quantikine ELISA Kit

  (4 citations)     
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Assay Procedure
Citations (4)
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  • 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 (100 uL), Serum (25 uL), EDTA Plasma (25 uL), Heparin Plasma (25 uL)
  • Sensitivity
    18.2 pg/mL
  • Assay Range
    46.9 - 3,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant porcine IL-12/IL-23 p40
  • 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.
Product Summary
The Quantikine Porcine IL-12/IL-23 p40 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure porcine IL-12/IL-23 p40 in cell culture supernates, serum and plasma. It contains CHO cell-expressed recombinant porcine IL-12 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural porcine IL-12/IL-23 p40 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring porcine 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
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 27 27 27
Mean 89.8 362 819 88.8 361 795
Standard Deviation 7.3 17.7 28 9.2 22.9 46.1
CV% 8.1 4.9 3.4 10.4 6.3 5.8

Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 25 25 25
Mean 115 447 991 110 454 995
Standard Deviation 7.4 22.7 33 10.1 33.4 64.3
CV% 6.4 5.1 3.3 9.2 7.4 6.5

Recovery

The recovery of porcine 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 97-113
EDTA Plasma (n=4) 92 84-99
Heparin Plasma (n=5) 94 85-105
Serum (n=5) 99 88-109
Linearity
To assess the linearity of the assay, samples spiked with or containing porcine IL-12/IL-23 p40 in each matrix were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
 IL-12/IL-23 p40 [HRP]
Product Datasheets

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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:
    CLMF2; IL12 p40; IL-12 p40; IL12B; IL-12B; NKSF2
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, 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. 100 µL Standard, Control, or Sample
  6.   Add 100 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 5 times.

  11. 120 µL Substrate Solution
  12.   Add 120 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 120 µL Stop Solution
  14.   Add 120 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.

4 Citations: Showing 1 - 4
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Species
Sample Type
  1. Dissecting the effects of lipopolysaccharides from nonlipopolysaccharide molecules in experimental porcine meningococcal sepsis.
    Authors: Hellerud BC, Nielsen EW, Thorgersen EB, Lindstad JK, Pharo A, Tonnessen TI, Castellheim A, Mollnes TE, Brandtzaeg P
    Crit. Care Med., 2010;38(6):1467-74.
    Species: Porcine
    Sample Type: Plasma
  2. CD14 inhibition efficiently attenuates early inflammatory and hemostatic responses in Escherichia coli sepsis in pigs.
    Authors: Thorgersen EB, Hellerud BC, Nielsen EW, Barratt-Due A, Fure H, Lindstad JK, Pharo A, Fosse E, Tonnessen TI, Johansen HT, Castellheim A, Mollnes TE
    FASEB J., 2010;24(3):712-22.
    Species: Porcine
    Sample Type: Plasma
  3. Metabolic activity of the enteric microbiota influences the fatty acid composition of murine and porcine liver and adipose tissues.
    Authors: Wall R, Ross RP, Shanahan F, O'Mahony L, O'Mahony C, Coakley M, Hart O, Lawlor P, Quigley EM, Kiely B, Fitzgerald GF, Stanton C
    Am. J. Clin. Nutr., 2009;89(5):1393-401.
    Species: Porcine
    Sample Type: Cell Culture Supernates
  4. Cytokine secretion depends on Galalpha(1,3)Gal expression in a pig-to-human whole blood model.
    Authors: Saethre M, Schneider MK, Lambris JD, Magotti P, Haraldsen G, Seebach JD, Mollnes TE
    J. Immunol., 2008;180(9):6346-53.
    Species: Porcine
    Sample Type: Cell Culture Supernates

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