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Human CXCL5/ENA-78 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB254

R&D Systems, part of Bio-Techne
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MAB254-100
MAB254-500
MAB254-SP

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunohistochemistry, Neutralization, Western Blot

Cited:

ELISA Development, Immunohistochemistry, Neutralization, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 33160

Product Specifications

Immunogen

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

Specificity

Detects human CXCL5/ENA‑78 in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant human (rh) CXCL1, 2, 3, 8, 10, or recombinant mouse CXCL2 is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Endotoxin Level

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

Scientific Data Images for Human CXCL5/ENA-78 Antibody

CXCL5/ENA-78 in Human Colon Cancer Tissue.

CXCL5/ENA-78 was detected in immersion fixed paraffin-embedded sections of human colon cancer tissue using Mouse Anti-Human CXCL5/ENA-78 Monoclonal Antibody (Catalog # MAB254) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in epithelial cells and secreted protein. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Chemotaxis Induced by CXCL5/ENA‑78 and Neutral-ization by Human CXCL5/ENA‑78 Antibody.

Chemotaxis Induced by CXCL5/ENA‑78 and Neutral-ization 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 Mouse Anti-Human CXCL5/ENA-78 Monoclonal Antibody (Catalog # MAB254). The ND50is typically 3-15 µg/mL.

Applications for Human CXCL5/ENA-78 Antibody

Application
Recommended Usage

Immunohistochemistry

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human colon cancer tissue

Western Blot

1 µg/mL
Sample: Recombinant Human CXCL5/ENA-78 (Catalog # 254-XB)
under non-reducing conditions only

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 3-15 µg/mL in the presence of 0.03 µg/mL Recombinant Human CXCL5/ENA-78.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from ascites

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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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.

Background: CXCL5/ENA-78

CXCL5, also known as epithelial cell-derived-neutrophil-activating-peptide (ENA78), 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. 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/GCP2 and 35%‑51% with CXCL1/GRO alpha, CXCL2/GRO beta, CXCL3/GRO gamma, CXCL7/NAP2, and CXCL8/IL8. 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. Its upregulation contributes to increased vascularization, tumor growth, and metastasis in many cancers. Full length CXCL5 (78 aa) is trimmed at the N-terminal end by Cathepsin G and chymotrypsin to ENA74 (74 aa) and ENA70 (70 aa), with the shortened forms showing increased potency relative to full length CXCL5. CXCL5 exerts its effects primarily through interactions with CXCR2. It also binds DARC, a decoy chemokine receptor which can limit CXCR2-mediated responses.

Alternate Names

CXCL5, ENA-78, ENA78

Entrez Gene IDs

6374 (Human)

Gene Symbol

CXCL5

UniProt

Additional CXCL5/ENA-78 Products

Product Documents for Human CXCL5/ENA-78 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human CXCL5/ENA-78 Antibody

For research use only

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