Human IL-5 R alpha/CD125 Antibody

Name: Human IL-5 R alpha/CD125 Antibody
Brand: R&D Systems
SKU: AF-253-NA

R&D Systems, part of Bio-Techne | Catalog # AF-253-NA

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Citations (4)

Product Datasheet / COA / SDS

View Available Conjugates

Conjugate

Catalog #

Biotin

BAF253

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

CyTOF-ready, Flow Cytometry, Neutralization, Western Blot

Cited:

Flow Cytometry, Immunocytochemistry, Neutralization, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

View all IL-5 R alpha/CD125 Antibodies »

Product Specifications

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant human IL‑5 R alpha/CD125

Specificity

Detects human IL-5 R alpha in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 5% cross-reactivity with recombinant human (rh) IL‑1 RII, rhIL-2 R beta, rhIL-2 R gamma, rhIL-3 R is observed and less than 1% cross-reactivity with rhIL‑1 RI, rhIL‑4 R, rhIL‑6 R, rhIL‑7 R, rhIL‑9 R, and rhIL‑10 R is observed.

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 IL-5 R alpha/CD125 Antibody

Cell Proliferation Induced by IL‑5 and Neutralization by Human IL‑5 R alpha/CD125 Antibody.

Recombinant Human IL-5 (205-IL) stimulates proliferation in the TF-1 human erythroleukemic cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human IL-5 (0.5 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human IL-5 Ra/CD125 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-253-NA). The ND₅₀ is typically 1-3 µg/mL.

Detection of IL-5 R alpha/CD125 in HEK293 human embryonic kidney cell line transfected with human IL-5 receptor alpha vs. HEK293 human embryonic kidney cell line transfected with an irrelevant protein by Flow Cytometry.

HEK293 human embryonic kidney cell line transfected with human IL-5 receptor alpha (filled histogram) vs HEK293 human embryonic kidney cell line transfected with an irrelevant protein (open histogram) were stained with Goat Anti-Human IL-5 R alpha/CD125 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-253-NA) followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). View our protocol for Staining Membrane-associated Proteins.

Applications for Human IL-5 R alpha/CD125 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.

Flow Cytometry

0.25 µg/10⁶ cells
Sample: HEK293 human embryonic kidney cell line transfected with human IL-5 receptor alpha vs HEK293 human embryonic kidney cell line transfected with an irrelevant protein.

Western Blot

0.1 µg/mL
Sample: Recombinant Human IL-5 R alpha/CD125 (Catalog # 253-5R)

Neutralization

Measured by its ability to neutralize IL-5-induced proliferation in the TF-1 human erythroleukemic cell line. Kitamura, T. et al. (1989) J. Cell Physiol. 140:323. The Neutralization Dose (ND₅₀) is typically 1-3 µg/mL in the presence of 0.5 ng/mL Recombinant Human IL-5.

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

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. See Certificate of Analysis for details.
*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: IL-5 R alpha/CD125

Interleukin 5, produced primarily by activated T cells and mast cells, has diverse biological effects on a variety of cell types. Human IL-5 is a potent eosinophil differentiation and activation factor in vivo and in vitro. Additionally, it has also been reported that IL-5 can stimulate the proliferation and/or differentiation of basophils and B cells. The multiple effects of IL-5 are mediated by binding of the cytokine to specific cell surface receptors expressed on target cells. As is the case with many other cytokines, the functional high-affinity receptor for IL-5 is a complex consisting of a ligand binding subunit ( alpha chain) and a second subunit ( beta chain) that can modulate the ligand binding affinity of the receptor complex. In the case of IL-5, the beta subunit is shared with the high affinity receptor complexes for IL-3 and GM-CSF. The beta chain does not bind any of the cytokines in question but is indispensable for the cytokine-mediated signaling.

cDNA clones for the alpha chain (IL-5 R alpha) of both the mouse and human high affinity IL-5 receptor complexes have been isolated. Human and mouse IL-5 R alpha are both members of the hematopoietin receptor superfamily characterized by the presence of the WSXWS, and a four cysteine residue motif in the extracellular domain of the transmembrane protein. In addition to the cDNA clone encoding the full-length transmembrane protein, cDNA clones that arise from alternative splicing and that encode soluble secreted forms of IL-5 R alpha have been isolated from mouse as well as human cells. A naturally-occurring soluble form of the IL-5 R alpha has been detected in biological fluids of autoimmune-prone mice and mice bearing chronic B cell leukemia (BCL₁).

A recombinant human IL-5 soluble receptor alpha has been shown to bind the human IL-5 dimer in a 1:1 ratio and acts as a human IL-5 antagonist. This molecule inhibits the proliferation of IL-5-dependent cell lines and blocks human umbilical cord blood eosinophil differentiation.