IL‑3 R alpha/CD123 was detect-ed in immersion fixed human peripheral blood lymphocytes using 2 µg/mL Mouse Anti-Human IL‑3 R alpha/CD123 Monoclonal Antibody (Catalog # MAB301) for 3 hours at room temperature. Cells were stained (red) and counter-stained (green). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

IL‑3 R alpha/CD123 in Human Tonsil.

IL-3 Ra/CD123 was detected in immersion fixed paraffin-embedded sections of human tonsil using 15 µg/mL Mouse Anti-Human IL-3 Ra/CD123 Monoclonal Antibody (Catalog # MAB301) overnight at 4 °C. Tissue was stained with the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; CTS002) and counter-stained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Cell Proliferation Induced by IL‑3 and Neutralization by Human IL‑3 R alpha/CD123 Antibody.

Recombinant Human IL-3 (203-IL) stimulates proliferation in the TF-1 human erythroleukemic cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human IL-3 (0.5 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human IL-3 Ra/CD123 Monoclonal Antibody (Catalog # MAB301). The ND₅₀ is typically 0.6-1.2 µg/mL.

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Applications for Human IL-3R alpha/CD123 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: THP-1 human acute monocytic leukemia cell line

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed human peripheral blood lymphocytes

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human tonsil

Western Blot

1 µg/mL
Sample: Recombinant Human IL‑3 R alpha/CD123 (Catalog # 301-R3) under non-reducing conditions only

Neutralization

Measured by its ability to neutralize IL‑3-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 0.6-1.2 µg/mL in the presence of 0.5 ng/mL Recombinant Human IL‑3.

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

Reviewed Applications

Read 4 reviews rated 4 using MAB301 in the following applications:

Immunohistochemistry (1 Review)
Microarray (1 Review)
Microarrays (2 Reviews)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

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: IL-3R alpha

IL-3 is a pleiotropic cytokine that can stimulate proliferation and differentiation of pluripotent hematopoietic stem cells as well as various lineage committed progenitors (1, 2). IL-3 exerts its activity through binding to a specific cell surface receptor known as IL-3 R. IL-3 R is a heterodimeric structure composed of a 70 kDa IL-3 R alpha subunit (CD123) and a 120-140 kDa IL-3 R beta subunit (CD131) (3, 4). IL-3 R alpha binds IL-3 with relatively low affinity. In the presence of IL-3 R beta, however, IL-3 R alpha has a much higher affinity for IL-3. It is not clear how signal transduction occurs following IL-3 binding. The IL-3 R alpha chain has a very short intracellular domain while the IL‑3 R beta chain has a very large cytoplasmic domain. The IL‑3 R beta chain is also shared by the receptors for IL-5 and GM-CSF. Cells known to express IL-3 receptors include hematopoietic progenitors, epithelial cells, double negative T cells, mast cells, basophils and blood monocytes (5).

References

Moore, M.A.S. et al. (1991) Blood 72:944.
Warren, D.J. et al. (1988) J. Immunol. 140:94.
Plant M. et al. (1989) Nature 339:150.
Budel, L.M. et al. (1990) Blood 75:1439.
Schrader, J.W. et al. (1988) In Interleukin-3: The Panspecific hemopoietin (ed. J.W. Schrader), Academic Press, San Diego, CA.

Long Name

Interleukin 3 Receptor alpha

Alternate Names

CD123, IL-3 R alpha, IL-3Ra, IL3R alpha, IL3RA

Entrez Gene IDs

3563 (Human); 16188 (Mouse); 102138639 (Cynomolgus Monkey)

Gene Symbol

IL3RA

UniProt

P26951

Additional IL-3R alpha Products

Product Documents for Human IL-3R alpha/CD123 Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Human IL-3R alpha/CD123 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human IL-3R alpha/CD123 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB301

Key Product Details

Species Reactivity

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Cited:

Applications

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Label

Antibody Source

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Product Specifications

Immunogen

Specificity

Clonality

Host

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Scientific Data Images for Human IL-3R alpha/CD123 Antibody

IL‑3 R alpha/CD123 in Human Peripheral Blood Lymphocytes.