Porcine IL-8/CXCL8 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY535
Ancillary Products Available
Porcine IL-8 / CXCL8 ELISA Standard Curve
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Product Details
Procedure
Citations (12)
FAQs
Supplemental Products
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Porcine IL-8/CXCL8 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant porcine CXCL8/IL-8. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

Normal Goat Serum: (Catalog # DY005)

 

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Normal Goat Serum: (Catalog # DY005)

 

Data Example

Porcine IL-8 / CXCL8 ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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; LUCT; LYNAP; MDNCF; MDNCFb-ENAP; member 8; MONAP; MONAPGCP1; NAF; NAP1; NAP-1NAP1; NCF; Neutrophil-activating protein 1; Protein 3-10C; T cell chemotactic factor; T-cell chemotactic factor; TCF; TSG1

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent with NGS, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Porcine IL-8/CXCL8 DuoSet 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.

12 Citations: Showing 1 - 10
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  1. Beneficial Effect of Mildly Pasteurized Whey Protein on Intestinal Integrity and Innate Defense in Preterm and Near-Term Piglets
    Authors: M Navis, V Muncan, PT Sangild, L Møller Wil, PJ Koelink, ME Wildenberg, E Abrahamse, T Thymann, RM van Elburg, IB Renes
    Nutrients, 2020;12(4):.
    Species: Porcine
    Sample Types: Tissue Homogenates
  2. Necrotizing enterocolitis is associated with acute brain responses in preterm pigs
    Authors: J Sun, X Pan, LI Christians, XL Yuan, K Skovgaard, DEW Chatterton, SS Kaalund, F Gao, PT Sangild, S Pankratova
    J Neuroinflammation, 2018;15(1):180.
    Species: Porcine
    Sample Types: Tissue Homogenates
  3. Characterization of blunt chest trauma in a long-term porcine model of severe multiple trauma
    Sci Rep, 2016;6(0):39659.
    Species: Porcine
    Sample Types: BALF
  4. Acute social stress-induced immunomodulation in pigs high and low responders to ACTH
    Authors: Blandine Lieubeau
    Physiol. Behav., 2016;169(0):1-8.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  5. Human milk oligosaccharide effects on intestinal function and inflammation after preterm birth in pigs
    Authors: Stine B Bering
    J. Nutr. Biochem., 2016;40(0):141-154.
    Species: Porcine
    Sample Types: Tissue Homogenates
  6. Local Epidermal Growth Factor Receptor Signaling Mediates the Systemic Pathogenic Effects of Staphylococcus aureus Toxic Shock Syndrome
    PLoS ONE, 2016;11(7):e0158969.
    Species: Porcine
    Sample Types: Vaginal Fluid
  7. Antimicrobial and immunomodulatory activities of PR-39 derived peptides.
    Authors: Veldhuizen E, Schneider V, Agustiandari H, van Dijk A, Tjeerdsma-van Bokhoven J, Bikker F, Haagsman H
    PLoS ONE, 2014;9(4):e95939.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  8. The immune response against Chlamydia suis genital tract infection partially protects against re-infection.
    Authors: De Clercq E, Devriendt B, Yin L, Chiers K, Cox E, Vanrompay D
    Vet Res, 2014;45(0):95.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  9. Characterization of homologous and heterologous adaptive immune responses in porcine reproductive and respiratory syndrome virus infection.
    Authors: Diaz I, Gimeno M, Darwich L, Navarro N, Kuzemtseva L, Lopez S, Galindo I, Segales J, Martin M, Pujols J, Mateu E
    Vet. Res., 2012;43(1):30.
    Species: Porcine
    Sample Types: Serum
  10. Cytokine protein expression levels in tracheobronchial lymph node homogenates of pigs infected with pseudorabies virus.
    Authors: Miller LC, Zanella EL, Waters WR
    Clin. Vaccine Immunol., 2010;17(5):728-34.
    Species: Porcine
    Sample Types: Tissue Homogenates
  11. Gas exchange and lung inflammation using nasal intermittent positive-pressure ventilation versus synchronized intermittent mandatory ventilation in piglets with saline lavage-induced lung injury: an observational study.
    Authors: Lampland AL, Meyers PA, Worwa CT, Swanson EC, Mammel MC
    Crit. Care Med., 2008;36(1):183-7.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  12. Impact of topical cooling solution and prediction of pulmonary graft viability from non-heart-beating donors.
    Authors: Inci I, Arni S, Inci D, Zhai W, Hillinger S, Leskosek B, Vogt P, Weder W
    J. Heart Lung Transplant., 2008;27(9):1016-22.
    Species: Porcine
    Sample Types: BALF

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