Porcine IL-10 Quantikine ELISA Kit

  (10 citations)     
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Assay Procedure
Citations (10)
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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 (50 uL), Serum (50 uL), EDTA Plasma (50 uL)
  • Sensitivity
    5.5 pg/mL
  • Assay Range
    31.3 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma)
  • Specificity
    Natural and recombinant porcine IL-10
  • 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-10 Immunoassay is a 4.5 hour solid phase ELISA designed to measure porcine IL-10 levels in cell culture supernates, porcine serum, and EDTA plasma. It contains E. coli-expressed recombinant porcine IL-10 and antibodies raised against the recombinant protein. Results obtained for naturally occurring porcine IL-10 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 of natural porcine IL-10.

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
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 88 264 791 89 264 802
Standard Deviation 3.7 6.8 27.5 6.4 11.8 54.7
CV% 4.2 2.6 3.5 7.2 4.5 6.8

Recovery

The recovery of porcine IL-10 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) 106 101-109
EDTA Plasma (n=4) 92 80-100
Serum (n=4) 94 84-102
Linearity
To assess the linearity of the assay, four samples spiked with high concentrations of porcine IL-10 in each matrix were diluted with Calibrator Diluent and then assayed.
 IL-10 [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-10
Interleukin-10 functions as an anti-inflammatory cytokine by inhibiting the expansion and activation of Th1 cells and Th17 cells and by promoting the development of M2 macrophages and regulatory T cells (Treg). Within a tumor microrenvironment, however, IL-10 can inhibit the expansion of both Treg and myeloid-derived suppressor cells (MDSC). IL-10 exerts protective effects including limiting tissue damage in arthritic inflammation and promoting muscle regeneration after injury, but it also contributes to the persistence of viral infections. IL-10 signals through a receptor complex composed of IL-10 R alpha and IL-10 R beta. IL-10 R beta additionally associates with IL-20 R alpha, IL-22 R alpha 1, or IL-28 R alpha to form the receptor complexes for IL-22, IL-26, IL-28, and IL-29.
  • Long Name:
    Interleukin 10
  • Entrez Gene IDs:
    3586 (Human); 16153 (Mouse); 25325 (Rat); 397106 (Porcine); 403628 (Canine); 493683 (Feline); 100135553 (Guinea Pig); 2949786 (Viral)
  • Alternate Names:
    CSIF; CSIFMGC126450; Cytokine synthesis inhibitory factor; IL10; IL-10; IL10A; IL-10MGC126451; interleukin 10; interleukin-10; TGIF
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. 100 µL Assay Diluent
  4.   Add 100 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.

10 Citations: Showing 1 - 10
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Species
Sample Type
  1. Butyrate modifies intestinal barrier function in IPEC-J2 cells through a selective upregulation of tight junction proteins and activation of the Akt signaling pathway
    Authors: H Yan, KM Ajuwon
    PLoS ONE, 2017;12(6):e0179586.
    Species: Porcine
    Sample Type: Cell Culture Supernates
  2. Effect of Lactobacillus salivarius bacteriocin Abp118 on the mouse and pig intestinal microbiota.
    Authors: Riboulet-Bisson E, Sturme MH, Jeffery IB
    PLoS ONE, 2012;7(2):e31113.
    Species: Porcine
    Sample Type: Serum
  3. Comparative measurement of cell-mediated immune responses of swine to the M and N proteins of porcine reproductive and respiratory syndrome virus.
    Authors: Jeong HJ, Song YJ, Lee SW
    Clin. Vaccine Immunol., 2010;17(4):503-12.
    Species: Porcine
    Sample Type: Cell Culture Supernates
  4. Inhibition of complement and CD14 attenuates the Escherichia coli-induced inflammatory response in porcine whole blood.
    Authors: Thorgersen EB, Pharo A, Haverson K, Axelsen AK, Gaustad P, Kotwal GJ, Sfyroera G, Mollnes TE
    Infect. Immun., 2009;77(2):725-32.
    Species: Porcine
    Sample Type: Whole Blood
  5. 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
  6. Blood concentrations of the cytokines IL-1beta, IL-6, IL-10, TNF-alpha and IFN-gamma during experimentally induced swine dysentery.
    Authors: Kruse R, Essen-Gustavsson B, Fossum C, Jensen-Waern M
    Acta Vet. Scand., 2008;50(0):32.
    Species: Porcine
    Sample Type: Serum
  7. 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
  8. Experimental severe Pseudomonas aeruginosa pneumonia and antibiotic therapy in piglets receiving mechanical ventilation.
    Authors: Luna CM, Baquero S, Gando S, Patron JR, Morato JG, Sibila O, Absi R, Famiglietti A, Vay CA, Von Stecher F, Agusti C, Torres A
    Chest, 2007;132(2):523-31.
    Species: Porcine
    Sample Type: Serum
  9. Butyrate differentially regulates cytokines and proliferation in porcine peripheral blood mononuclear cells.
    Authors: Weber TE, Kerr BJ
    Vet. Immunol. Immunopathol., 2006;113(1):139-47.
    Species: Porcine
    Sample Type: Cell Culture Supernates
  10. Conjugated linoleic acid attenuates the production and gene expression of proinflammatory cytokines in weaned pigs challenged with lipopolysaccharide.
    Authors: Changhua L, Jindong Y, Defa L, Lidan Z, Shiyan Q, Jianjun X
    J. Nutr., 2005;135(2):239-44.
    Species: Porcine
    Sample Type: Serum

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