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Canine IL-8/CXCL8

Canine IL-8/CXCL8 Quantikine ELISA Kit Summary

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.

Preparation and Storage

Shipping

The   product is shipped at ambient temperature. Upon receipt, store it immediately   at the temperature recommended below.

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

Refer to the product datasheet for the 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.

1Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3Add 50 μL of Assay Diluent to each well.

4Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7Aspirate and wash 5 times.

8Add 100 μL Substrate Solution to each well.

9Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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