Canine IL-8/CXCL8 Antibody

Catalog #: MAB1608 Datasheet

Catalog #	Availability	Size / Price	Qty
MAB1608
MAB1608-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 canine CXCL8/IL-8 in direct ELISAs and Western blots. Does not cross-react with recombinant human CXCL1, 2, 3, 5, 6, 7, 8, 9, 10, 11, CXCL12/SDF-1 alpha, 12/SDF-1 beta, 13, 16, recombinant mouse CXCL1, 6, 9, 10, CXCL12/SDF-1 alpha, 13, recombinant rat (rr) CXCL1, CXCL3/CINC-2 alpha, rrCXCL3/CINC-2 beta, or recombinant porcine CXCL8.

Source

Monoclonal Mouse IgG₁ Clone # 258911

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived recombinant canine CXCL8/IL-8
Ala23-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

1 µg/mL

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

Immunocytochemistry

8-25 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 0.3-1.2 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 Mouse Anti-Canine IL-8/CXCL8 Monoclonal Antibody (Catalog # MAB1608). The ND₅₀ is typically 0.3-1.2 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 calcium ionomycin and PMA using Mouse Anti-Canine IL-8/CXCL8 Monoclonal Antibody (Catalog # MAB1608) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Preparation and Storage

Reconstitution

Reconstitute at 0.5 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