Canine IL-8/CXCL8 Antibody

Catalog #: AF1608 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF1608
AF1608-SP

Chemotaxis Induced by IL-8/CXCL8 and Neutralization by Canine IL-8/CXCL8 Antibody.

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Canine IL-8/CXCL8 Antibody Summary

Species Reactivity

Canine

Specificity

Detects IL-8/CXCL8 in direct ELISAs and Western blots. In Western blots, less than 1% cross-reactivity with recombinant human IL-8/CXCL8, recombinant porcine IL-8/CXCL8, recombinant rat (rr) CINC-1, rrCINC-2 alpha, and rrCINC-2 beta is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant canine IL-8/CXCL8
Ala23-Pro101 & & Val28-Pro101
Accession # P41324

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

Recombinant Canine IL-8/CXCL8 (Catalog # 1608-CL)

Immunocytochemistry

5-15 µg/mL

See below

Neutralization

Measured by its ability to neutralize IL-8/CXCL8-induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with human CXCR2. The Neutralization Dose (ND₅₀) is typically 0.1-0.5 µg/mL in the presence of 10 ng/mL Recombinant Canine IL-8/CXCL8.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Neutralization

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Chemotaxis Induced by IL-8/CXCL8 and Neutralization by Canine IL-8/CXCL8 Antibody. Recombinant Canine IL-8/CXCL8 (Catalog # 1608-CL) chemoattracts the BaF3 mouse pro-B cell line transfected with human CXCR2 in a dose-dependent manner (orange line). The amount of cells that migrated through to the lower chemotaxis chamber was measured by Resazurin (Catalog # AR002). Chemotaxis elicited by Recombinant Canine IL-8/CXCL8 (10 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Canine IL-8/CXCL8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1608). The ND₅₀ is typically 0.1-0.5 µg/mL.

Immunocytochemistry

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IL-8/CXCL8 in Canine PBMCs. IL-8/CXCL8 was detected in immersion fixed canine peripheral blood mononuclear cells (PBMCs) treated with Concanavalin A using Goat Anti-Canine IL-8/CXCL8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1608) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: IL-8/CXCL8

Interleukin 8 (IL-8), also named monocyte-derived neutrophil chemotactic factor (MDNCF), neutrophil-activating protein 1 (NAP-1), neutrophil-activating factor (NAF) and granulocyte chemotactic peptide (GCP), belongs to the Glu-Leu-Arg motif containing (ELR⁺) CXC chemokine family and has been designated CXCL8. IL-8 is a potent neutrophil chemoattractant that recruits neutrophils to sites of inflammation. IL-8 also activates neutrophil functions and through a poorly understood mechanism, promotes angiogenesis. The biological activites of IL-8 is mediated by two types of G protein-coupled chemokine receptors, CXCR1 and CXCR2. In normal tissues, IL-8 expression and secretion is barely detectable. Upon stimulation by a wide range of pro-inflammatory signals including exposure to IL-1, TNF, bacterial or viral products, IL-8 production is rapidly induced in many different cell types. Secreted IL-8 is not glycosylated but has N‑terminal sequence heterogenecity due to proteolytic processing. In human, two major forms, the 72 amino acid (aa) monocyte-derived IL-8 and the 77 aa endothelial IL-8 have been identified. Whereas the 72 aa isoform is a more potent chemoattractant, only the 77 aa isoform can induce apoptosis in leukemic cells. The N‑terminal pentapeptide in the 77 aa isoform has been identified as the active site for the IL-8 apoptotic activity. Canine IL-8 encodes a 101 aa precursor protein with a putative 22 aa signal peptide. It shares 77% and 87% aa sequence identity with human and porcine IL-8, respectively. Similar to human IL-8, recombinant canine IL-8 also undergoes
N‑terminal processing. Two major peptides (the 79 aa and 74 aa variants that differ by an analogous N‑terminal pentapeptide) are present in the recombinant canine IL-8 preparations.

References

Van Damme, J. et al. (1998) in The Cytokine Handbook, A.W. Thomson, ed., Academic Press, New York., p. 271.
Terui, Y. et al. (1998) Blood, 92:2672.
Terui, Y. et al. (1999) Cancer Research 59:5651.

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