Human CXCL5/ENA-78 Antibody AF254: R&D Systems

Human CXCL5/ENA-78 Antibody

(5 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human CXCL5/ENA‑78 in direct ELISAs and Western blots. In direct ELISAs, approximately 20% cross-reactivity with recombinant human (rh) GCP-2 is observed and less than 5% cross-reactivity with recombinant mouse (rm) KC, recombinant rat CINC-1, rhGRO alpha, rhGRO gamma and rmGCP-2 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human CXCL5/ENA-78
    Ala37-Asn114
    Accession # P42830
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied 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 Human CXCL5/ENA‑78 (Catalog # 254-XB)
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize CXCL5/ENA‑78-induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with human CXCR2. The Neutralization Dose (ND50) is typically 0.2-1.0 µg/mL in the presence of 0.03 µg/mL Recombinant Human CXCL5/ENA‑78.
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.
Data Examples
Chemotaxis Induced by CXCL5/ENA‑78 and Neutralization by Human CXCL5/ENA‑78 Antibody. Recombinant Human CXCL5/ENA‑78 (Catalog # 254‑XB) 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 Human CXCL5/ENA‑78 (0.03 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human CXCL5/ENA‑78 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF254). The ND50 is typically 0.2-1.0 µg/mL.
Immunohistochemistry
CXCL5/ENA‑78 in Human Breast. CXCL5/ENA‑78 was detected in immersion fixed paraffin-embedded sections of human breast using Goat Anti-Human
CXCL5/ENA‑78 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF254) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Immunohistochemistry
CXCL5/ENA‑78 in Human Breast Cancer Tissue. CXCL5/ENA‑78 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Goat Anti-Human CXCL5/ENA‑78 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF254) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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: CXCL5/ENA-78

CXCL5, also known as epithelial cell-derived neutrophil-activating peptide (ENA-78), is an 8 kDa proinflammatory member of the CXC subfamily of chemokines. Its Glu-Leu-Arg (ELR) motif confers angiogenic properties and distinguishes it from ELR-CXC chemokines which are angiostatic (1‑3). Human CXCL5 shares 57% amino acid (aa) sequence identity with mouse and rat CXCL5. Among other human ELR+ chemokines, it shares 77% aa sequence identity with CXCL6/GCP-2 and
35%‑51% with CXCL1/GRO alpha, CXCL2/GRO beta, CXCL3/GRO gamma, CXCL7/NAP-2, and CXCL8/IL-8. Inflammatory stimulation upregulates CXCL5 production in multiple hematopoietic cell types, fibroblasts, endothelial cells, and vascular smooth muscle cells. In vivo, CXCL5 is elevated at sites of inflammation and pulmonary fibrosis where it promotes neutrophil infiltration and activation as well as angiogenesis (3 - 6). Its upregulation contributes to increased vascularization, tumor growth, and metastasis in many cancers (6 - 9). Full length CXCL5 (78 aa) is trimmed at the N-terminus by cathepsin G and chymotrypsin to ENA-74 (74 aa) and ENA-70 (70 aa), with the shortened forms showing increased potency relative to full length CXCL5 (10, 11). CXCL5 exerts its effects primarily through interactions with CXCR2 (6, 12). It also binds duffy antigen receptor for chemokines (DARC), which can limit CXCR2-mediated responses (13, 14).

  • References:
    1. Strieter, R.M. et al. (2005) Cytokine Growth Factor Rev. 16:593.
    2. Balestrieri, M.L. et al. (2008) Cardiovasc. Res. 78:250.
    3. Walz, A.. et al. (1991) J. Exp. Med. 174:1355.
    4. Strieter, R.M. et al. (1992) Immunol. Invest. 21:549.
    5. Koch, A.E. et al. (1994) J. Clin. Invest. 94:1012.
    6. Begley, L.A. et al. (2008) Neoplasia 10:244.
    7. Vandercappellen, J. et al. (2008) Cancer Lett. 267:226.
    8. Arenberg, D.A. et al. (1998) J. Clin. Invest. 102:465.
    9. Miyazaki, H. et al. (2006) Cancer Res. 66:4279.
    10. Wuyts, A. et al. (1999) Eur. J. Biochem. 260:421.
    11. Nufer, O. et al. (1999) Biochemistry 38:636.
    12. Ahuja, S.K. and P.M. Murphy (1996) J. Biol. Chem. 271:20545.
    13. Kashiwazaki, M. et al. (2003) Int. Immunol. 15:1219.
    14. Horton, L.W. et al. (2007) Cancer Res. 67:9791.
  • Entrez Gene IDs:
    6374 (Human)
  • Alternate Names:
    CXCL5; CXCL5/ENA-78; ENA78; ENA-78
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

5 Citations: Showing 1 - 5
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Species
Applications
Sample Type
  1. CXCR2-mediated tumor-associated neutrophil recruitment is regulated by IFN-beta.
    Authors: Jablonska J, Wu C, Andzinski L, Leschner S, Weiss S
    Int J Cancer, 2014;134(6):1346-58.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC- Frozen
  2. Overexpression of CXCL5 is associated with poor survival in patients with pancreatic cancer.
    Authors: Li A, King J, Moro A, Sugi MD, Dawson DW, Kaplan J, Li G, Lu X, Strieter RM, Burdick M, Go VL, Reber HA, Eibl G, Hines OJ
    Am. J. Pathol., 2011;178(3):1340-9.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  3. A CXCL5- and bFGF-dependent effect of PDGF-B-activated fibroblasts in promoting trafficking and differentiation of bone marrow-derived mesenchymal stem cells.
    Authors: Nedeau AE, Bauer RJ, Gallagher K, Chen H, Liu ZJ, Velazquez OC
    Exp. Cell Res., 2008;314(11):2176-86.
    Species: Mouse
    Sample Type: Whole Cells
    Application: Neut
  4. Expression and gene polymorphisms of the chemokine CXCL5 in colorectal cancer patients.
    Authors: Dimberg J, Dienus O, Lofgren S, Hugander A, Wagsater D
    Int. J. Oncol., 2007;31(1):97-102.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Frozen
  5. Imbalance in the expression of CXC chemokines correlates with bronchoalveolar lavage fluid angiogenic activity and procollagen levels in acute respiratory distress syndrome.
    Authors: Keane MP, Donnelly SC, Belperio JA, Goodman RB, Dy M, Burdick MD, Fishbein MC, Strieter RM
    J. Immunol., 2002;169(11):6515-21.
    Species: Human
    Sample Type: BALF
    Application: ELISA Development
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