Canine IL-8/CXCL8 Quantikine ELISA Kit

(2 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 (50 uL), Serum (13 uL), EDTA Plasma (13 uL), Heparin Plasma (13 uL)
  • Sensitivity
    4.31 pg/mL
  • Assay Range
    15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant canine IL-8. This assay also recognizes the 74 and 79 amino acid variants of recombinant canine IL-8.
  • 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 Canine IL-8 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure canine IL-8 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant canine IL-8 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural canine IL-8 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 canine IL-8.

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
n202020393739
Mean38.810562843.9116682
Standard Deviation2.355.832.42.65.536.1
CV%65.65.25.94.75.3

Recovery

The recovery of canine IL-8 spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=6) 97 88-103
EDTA Plasma (n=4) 100 93-108
Heparin Plasma (n=4) 102 91-113
Serum (n=4) 97 91-101
Linearity
To assess the linearity of the assay, samples containing or spiked with various concentrations of canine IL-8 in each matrix were diluted with Calibrator Diluent and then assayed.
Canine IL-8/CXCL8 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: IL-8/CXCL8
Interleukin-8 (IL-8), also known as CXCL8, is a chemokine that is upregulated at sites of inflammation where it promotes neutrophil infiltration and activation. IL-8 can form homodimers and heterodimers with CXCL4/PF4. Its bioactivity is regulated by proteolytic truncations, citrullination, and the decoy receptor DARC. IL-8 signals through CXCR1/IL-8 RA, which is also used by CXCL6, and through CXCR2/IL-8 RB, which is used by multiple CXC chemokines. IL-8 also binds to Serpin A1/alpha-1 Antitrypsin which prevents IL-8 interaction with CXCR1.
    • Long Name
      Interleukin 8
    • Entrez Gene IDs
      3576 (Human); 396880 (Porcine); 403850 (Canine); 493836 (Feline);
    • Alternate Names
      3-10C; AMCF-I; C-X-C motif chemokine 8; CXCL8; CXCL8SCYB8; Emoctakin; GCP1; GCP-1TSG-1; IL8; IL-8; interleukin 8; K60; LAI; LECT; MDNCF; MDNCFb-ENAP; member 8; MONAPGCP1; NAP1; NAP-1NAP1; NCF; Neutrophil-activating protein 1; Protein 3-10C; T cell chemotactic factor; T-cell chemotactic factor; TCF; TSG1;
    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. Tap the plate gently for 1 minute. 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.

    2 Citations: Showing 1 - 2
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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. Continuous venovenous hemofiltration decreases mortality and ameliorates acute lung injury in canine model of severe salt water drowning
      Authors: J Chen, G Chen, D Xiao, W Peng, G Yu, Y Lin, F Zheng
      Scand J Trauma Resusc Emerg Med, 2016;24(1):40.
      Species: Canine
      Sample Type: Plasma
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    Average Rating: 4 (Based on 1 review)

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     Canine CXCL8/IL-8 Quantikine ELISA Kit CA8000
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      Anonymous 10/14/2016
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     Canine CXCL8/IL-8 Quantikine ELISA Kit CA8000
    : Canine CXCL8/IL-8 Quantikine ELISA Kit [CA8000]

    Summary

    Sample TestedCell Culture Supernates

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