Canine IL-10 Quantikine ELISA Kit

(6 citations)
(1 Review)
  
  • 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 (25 uL), Serum (25 uL), EDTA Plasma (25 uL), Heparin Plasma (25 uL)
  • Sensitivity
    3.8 pg/mL
  • Assay Range
    15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant canine IL-10
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    Interference observed with 1 or more available related molecules.
Product Summary
The Quantikine Canine IL-10 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure canine IL-10 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant canine IL-10 and antibodies raised against the recombinant protein. Results obtained for naturally occurring canine 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 for natural canine IL-10.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times 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
Sample123123
n202020202020
Mean5314453359152512
Standard Deviation2.9720.15.19.321
CV%5.54.93.88.66.14.1

Recovery

The recovery of canine IL-10 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=5) 93 75-104
EDTA Plasma (n=4) 89 83-93
Heparin Plasma (n=4) 87 83-93
Serum (n=5) 96 80-107
Linearity
To assess the linearity of the assay, samples containing or spiked with high concentrations of canine IL-10 in each matrix were diluted with Calibrator Diluent RD6-12 and assayed.
Canine IL-10 Quantikine ELISA Kit
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. 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. Gently tap the plate to ensure thorough mixing. 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.

    6 Citations: Showing 1 - 6
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    Species
    Sample Type
    1. Higher neonatal growth rate and body condition score at 7�months are predictive factors of obesity in adult female Beagle dogs
      Authors: L Leclerc, C Thorin, J Flanagan, V Biourge, S Serisier, P Nguyen
      BMC Vet. Res., 2017;13(1):104.
      Species: Canine
      Sample Type: Plasma
    2. Use of a recombinant cysteine proteinase from Leishmania (Leishmania) infantum chagasi for the Immunotherapy of canine visceral Leishmaniasis.
      Authors: Ferreira J, Silva L, Longo-Maugeri I, Katz S, Barbieri C
      PLoS Negl Trop Dis, 2014;8(3):e2729.
      Species: Canine
      Sample Type: Serum
    3. Programmed death 1-mediated T cell exhaustion during visceral leishmaniasis impairs phagocyte function.
      Authors: Esch K, Juelsgaard R, Martinez P, Jones D, Petersen C
      J Immunol, 2013;191(11):5542-50.
      Species: Canine
      Sample Type: Cell Culture Supernates
    4. Clinical, parasitologic, and immunologic evolution in dogs experimentally infected with sand fly-derived Leishmania chagasi promastigotes.
      Authors: Travi BL, Osorio EY, Saldarriaga OA
      Am. J. Trop. Med. Hyg., 2009;81(6):994-1003.
      Species: Canine
      Sample Type: Cell Culture Supernates
    5. Measurement of serum interleukin-10 in the dog.
      Authors: Kjelgaard-Hansen M, Luntang-Jensen M, Willesen J, Jensen AL
      Vet. J., 2007;173(2):361-5.
      Species: Canine
      Sample Type: Serum
    6. A functional comparison of canine and murine bone marrow derived cultured mast cells.
      Authors: Lin TY, London CA,
      Vet. Immunol. Immunopathol., 2006;114(3):320-34.
      Species: Canine
      Sample Type: Cell Culture Supernates
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    Average Rating: 5 (Based on 1 review)

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    Summary

    Sample Testedcanine serum samples,Serum and Plasma

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