Human IL-8/CXCL8 Antibody

Catalog #: MAB2083 Datasheet

Catalog #	Availability	Size / Price	Qty
MAB2083-100
MAB2083-SP

Detection of Human IL-8/CXCL8 by Western Blot.

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Human IL-8/CXCL8 Antibody Summary

Species Reactivity

Human

Specificity

Detects human IL-8/CXCL8 in direct ELISAs.

Source

Monoclonal Mouse IgG₁ Clone # 1028336

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived human IL-8/CXCL8
Ser28-Ser99
Accession # P10145.1

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

3 µg/mL

THP‑1 human acute monocytic leukemia cell line treated with LPS and PMA

Simple Western

50 µg/mL

PBMC conditioned media treated with PMA, ionomycin, and LPS

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

Western Blot

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Detection of Human IL-8/CXCL8 by Western Blot. Western blot shows lysates of THP‑1 human acute monocytic leukemia cell line untreated (-) or treated (+) with 200 nM PMA for 24 hours and 10 µg/mL LPS for 3 hours. PVDF membrane was probed with 3 µg/mL of Mouse Anti-Human IL-8/CXCL8 Monoclonal Antibody (Catalog # MAB2083) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for IL-8/CXCL8 at approximately 11 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Simple Western

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Detection of Human IL-8/CXCL8 by Simple Western^TM. Simple Western lane view shows PBMC conditioned media untreated (-) or treated (+) with 2 µg/mL LPS for 24 hours, 200 ng/mL PMA for 24 hours, and 10 µg/mL ionomycin for 3 hours, loaded at 0.2 mg/mL. A specific band was detected for IL-8/CXCL8 at approximately 9 kDa (as indicated) using 50 µg/mL of Mouse Anti-Human IL-8/CXCL8 Monoclonal Antibody (Catalog # MAB2083). This experiment was conducted under reducing conditions and using the 2-40 kDa separation system.

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 known as CXCL8, GCP-1, and NAP-1, is a widely expressed proinflammatory member of the CXC family of chemokines. Near its N-terminus, this 8-9 kDa chemokine contains an ELR motif which is important for its angiogenic properties (1). IL-8/CXCL8 can associate into a homodimer or a heterodimer with CXCL4/PF4 (2), and it can also interact with matrix and cell surface glycosaminoglycans (3). Mature human IL-8/CXCL8 shares 65%-69% amino acid (aa) sequence identiity with canine, feline, and porcine IL-8/CXCL8 (4). There is no IL-8/CXCL8 gene counterpart in rodent. N-terminal truncation by multiple proteases generates a range of shorter forms, and an alternative splice form of human IL-8/CXCL8 carries an eleven aa substitution at the C-terminus (5). The bioactivity of IL-8/CXCL8 is regulated by these truncations, by IL-8/CXCL8 citrullination at Arg5 (N-terminal to the ELR motif) (6), and by the decoy receptor DARC (7). IL-8/CXCL8 effects are mediated through CXCR1/IL-8 RA, which is also used by CXCL6, and through CXCR2/IL-8 RB, which is used by multiple CXC chemokines (1). CXCR1 and CXCR2 associate into functional homodimers and heterodimers with each other (8). Through both CXCR1 and CXCR2, CXCL8 promotes neutrophil adhesion to the vascular endothelium and migration to sites of inflammation (9). It triggers the antimicrobial activation of neutrophils through CXCR1 (10). CXCL8 also binds to Serpin A1/alpha-1 Antitrypsin, and this prevents IL-8/CXCL8 interaction with CXCR1 (11). IL-8/CXCL8 is upregulated in atherosclerotic lesions and other cardiac pathologies where it exacerbates inflammatory tissue damage (12). In addition, it induces VEGF expression, vascular endothelial cell proliferation, angiogenesis, and tumor cell invasiveness (13-16).

References

Lazennec, G. and A. Richmond (2010) Trends Mol. Med. 16:133.
Nesmelova, I.V. et al. (2005) J. Biol. Chem. 280:4948.
Pichert, A. et al. (2012) Biomatter 2:142.
Schmid, J. and C. Weissmann (1987) J. Immunol. 139:250.
Mortier, A. et al. (2008) Pharmacol. Ther. 120:197.
Proost, P. et al. (2008) J. Exp. Med. 205:2085.
Neote, K. et al. (1994) Blood 84:44.
Munoz, L.M. et al. (2009) J. Immunol. 183:7337.
Gerszten, R.E. et al. (1999) Nature 398:718.
Jones, S.A. et al. (1996) Proc. Natl. Acad. Sci. USA 93:6682.
Bergin, D.A. et al. (2010) J. Clin. Invest. 120:4236.
Apostolakis, S. et al. (2009) Cardiovasc. Res. 84:353.
Martin, D. et al. (2009) J. Biol. Chem. 284:6038.
Li, A. et al. (2005) Angiogenesis 8:63.
Waugh, D.J. and C. Wilson (2008) Clin. Cancer Res. 14:6735.
Fernando, R.I. et al. (2011) Cancer Res. 71:5296.

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