Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Neutralization

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Neutralization, Flow Cytometry, ELISA Development, In vivo assay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all CXCL5/ENA-78 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human CXCL5/ENA-78
Ala37-Asn114
Accession # P42830

Specificity

Detects human CXCL5/ENA‑78 in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

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

Scientific Data Images for Human CXCL5/ENA‑78 Antibody

Detection of Human CXCL5/ENA-78 antibody by Western Blot.

Detection of Human CXCL5/ENA‑78 by Western Blot.

Western blot shows conditioned media (CM) and lysates from A549 human lung carcinoma cell line untreated (-) or treated (+) with human IL-1 beta for 24 hours. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human CXCL5/ENA-78 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF254) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). A specific band was detected for CXCL5/ENA-78 at approximately 10 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

CXCL5/ENA-78 antibody in Human Breast Cancer Tissue by Immunohistochemistry (IHC-P).

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; (CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

CXCL5/ENA-78 antibody in Human Breast by Immunohistochemistry (IHC-P).

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; (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.

Chemotaxis Induced by CXCL5/ENA‑78 and Neutralization by Human CXCL5/ENA‑78 Antibody.

Chemotaxis Induced by CXCL5/ENA‑78 and Neutralization by Human CXCL5/ENA‑78 Antibody.

Recombinant Human CXCL5/ENA-78 (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 (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.

Applications for Human CXCL5/ENA‑78 Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample:  Immersion fixed paraffin-embedded sections of human breast and human breast cancer tissue

Western Blot

1 µg/mL
Sample: Conditioned media and lysates from A549 human lung carcinoma cell line treated with human IL-1 beta

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.

Reviewed Applications

Read 3 reviews rated 4.7 using AF254 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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 up-regulates 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.

Alternate Names

CXCL5, ENA-78, ENA78

Entrez Gene IDs

6374 (Human)

Gene Symbol

CXCL5

UniProt

P42830

Additional CXCL5/ENA-78 Products

Product Documents for Human CXCL5/ENA‑78 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human CXCL5/ENA‑78 Antibody

For research use only

Citations for Human CXCL5/ENA‑78 Antibody

Customer Reviews for Human CXCL5/ENA‑78 Antibody (3)

4.7 out of 5
3 Customer Ratings
5 Stars
67%
4 Stars
33%
3 Stars
0%
2 Stars
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1 Stars
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Showing  1 - 3 of 3 reviews Showing All
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  • Human CXCL5/ENA-78 Antibody
    Name: Anonymous
    Application: Immunohistochemistry-Paraffin
    Sample Tested: Breast cancer tissue
    Species: Human
    Verified Customer | Posted 04/04/2020
    Human CXCL5/ENA‑78 Antibody AF254
  • Human CXCL5/ENA-78 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Hep3B human hepatocellular carcinoma cell line and SK-HEP-1 human liver adenocarcinoma cell line
    Species: Human
    Verified Customer | Posted 02/18/2019
    Human CXCL5/ENA‑78 Antibody AF254
  • Human CXCL5/ENA-78 Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Cancer Tissue
    Species: Human
    Verified Customer | Posted 10/17/2018
    Human CXCL5/ENA‑78 Antibody AF254

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Protocols

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