Human peripheral blood mononculear cells (PBMCs) either treated with LPS (filled histogram) or untreated (open histogram) were stained with Mouse Anti-Human CXCL5/ENA-78 Monoclonal Antibody (Catalog # MAB654), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

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 (30 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human CXCL5/ENA-78 Monoclonal Antibody (Catalog # MAB654). The ND₅₀is typically 0.65-5 µg/mL.

Human CXCL5 / ENA-78 ELISA Standard Curve

Recombinant Human CXCL5/ENA-78 (Catalog # 254-XB) was serially diluted and captured by Mouse Anti-Human CXCL5/ENA-78 Monoclonal Antibody (Catalog # MAB654) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human CXCL5/ENA-78 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF254) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

Applications for Human CXCL5/ENA-78 Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells
Sample: Human peripheral blood mononculear cells (PBMCs) treated with LPS were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005)

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 0.65-5 µg/mL in the presence of 30 ng/mL Recombinant Human CXCL5/ENA‑78.

Human CXCL5/ENA-78 Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)

Recommended Concentration: 2-8 µg/mL
Use in combination with these reagents:

Detection Reagent: Human CXCL5/ENA-78 Biotinylated Antibody (Catalog # BAF254)
Standard: Recombinant Human CXCL5/ENA-78 Protein (Catalog # 254-XB)

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 4 using MAB654 in the following applications:

ELISA (1 Review)

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.

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 ENA-78, is a CXC chemokine that plays a role in inflammatory responses and angiogenesis.

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

Product Datasheets

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COA

SDS

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Product Specific Notices for Human CXCL5/ENA-78 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human CXCL5/ENA-78 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB654

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications

Validated:

Cited:

Label

Antibody Source

View all CXCL5/ENA-78 Antibodies »

Product Specifications

Immunogen

Specificity

Clonality

Host

Isotype

Endotoxin Level

Scientific Data Images for Human CXCL5/ENA-78 Antibody

Detection of CXCL5/ENA‑78 in Human PBMCs by Flow Cytometry.