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.
Canine IL-8/CXCL8 Quantikine ELISA Kit
R&D Systems | Catalog # CA8000
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (13 µL), EDTA Plasma (13 µL), Heparin Plasma (13 µL)
Sensitivity
4.31 pg/mL
Assay Range
15.6-1000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Product Summary for Canine IL-8/CXCL8 Quantikine ELISA Kit
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.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Canine
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.
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, EDTA Plasma, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 39 | 37 | 39 |
| Mean (pg/mL) | 38.8 | 105 | 628 | 43.9 | 116 | 682 |
| Standard Deviation | 2.35 | 5.8 | 32.4 | 2.60 | 5.5 | 36.1 |
| CV% | 6.0 | 5.6 | 5.2 | 5.9 | 4.7 | 5.3 |
Recovery for Canine IL-8/CXCL8 Quantikine ELISA Kit
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.
Scientific Data Images for Canine IL-8/CXCL8 Quantikine ELISA Kit
Canine CXCL8/IL-8 ELISA Standard Curve
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
Long Name
Interleukin 8
Alternate Names
CXCL8, GCP1, IL8, LAI, LECT, LUCT, LYNAP, MDNCF, MONAP, NAF, NAP1, NCF, TCF, TSG1
Gene Symbol
CXCL8
Additional IL-8/CXCL8 Products
Product Documents for Canine IL-8/CXCL8 Quantikine ELISA Kit
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Canine IL-8/CXCL8 Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Citations for Canine IL-8/CXCL8 Quantikine ELISA Kit
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Protocols
View specific protocols for Canine IL-8/CXCL8 Quantikine ELISA Kit (CA8000):
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.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 50 µL of Assay Diluent to each well.
- 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.
- Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
- Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 5 times.
- Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- 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.
50 µL Assay Diluent
50 µL Standard, Control, or sample
100 µL Conjugate
100 µL Substrate Solution
100 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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