Human IL-3 R alpha /CD123 Antibody

(3 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human IL‑3 R alpha /CD123 in direct ELISAs and Western blots.
  • Source
    Monoclonal Mouse IgG1 Clone # 32703
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    S. frugiperda insect ovarian cell line Sf 21-derived recombinant human IL‑3 R alpha /CD123
    Lys20-Arg305, predicted
    Accession # P26951
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    Recombinant Human IL‑3 R alpha /CD123 (Catalog # 301-R3) under non-reducing conditions only
  • Flow Cytometry
    2.5 µg/106 cells
    THP‑1 human acute monocytic leukemia cell line
  • Immunohistochemistry
    8-25 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    8-25 µg/mL
    See below
  • 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 should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Immunocytochemistry
IL‑3 R alpha /CD123 in Human Peripheral Blood Lymphocytes. IL‑3 R alpha /CD123 was detect­ed 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 counter­stained (green). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Immunohistochemistry
IL‑3 R alpha /CD123 in Human Tonsil.
IL‑3 R alpha /CD123 was detected in immersion fixed paraffin-embedded sections of human tonsil using 15 µg/mL Mouse Anti-Human
IL‑3 R alpha /CD123 Monoclonal Antibody (Catalog # MAB301) overnight at 4 °C. Tissue was stained with the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counter­stained 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 (Catalog # 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 R alpha /CD123 Monoclonal Antibody (Catalog # MAB301). The ND50 is typically
0.6‑1.2 µg/mL.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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-3 R alpha/CD123
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:
    1. Moore, M.A.S. et al. (1991) Blood 72:944.
    2. Warren, D.J. et al. (1988) J. Immunol. 140:94.
    3. Plant M. et al. (1989) Nature 339:150.
    4. Budel, L.M. et al. (1990) Blood 75:1439.
    5. 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
  • Entrez Gene IDs:
    3563 (Human); 16188 (Mouse)
  • Alternate Names:
    CD123 antigen; CD123; hIL3Ra; hIL-3Ra; IL-3 R alpha; IL-3 receptor subunit alpha; IL3R alpha; IL-3R subunit alpha; IL3R; IL3RA; IL-3Ra; IL-3R-alpha; IL3RAY; IL3RX; IL3RY; interleukin 3 receptor, alpha (low affinity); interleukin-3 receptor subunit alpha; MGC34174
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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Species
Applications
Sample Type
  1. Interleukin-3 enhances the migration of human mesenchymal stem cells by regulating expression of CXCR4
    Authors: A Barhanpurk, ST Mhaske, ST Pote, K Singh, MR Wani
    Stem Cell Res Ther, 2017;8(1):168.
    Species: Human
    Sample Type: Whole Cells
    Application: Flow
  2. Graves' disease is associated with a defective expression of the immune regulatory molecule galectin-9 in antigen-presenting dendritic cells.
    Authors: Leskela S, Serrano A, de la Fuente H, Rodriguez-Munoz A, Ramos-Levi A, Sampedro-Nunez M, Sanchez-Madrid F, Gonzalez-Amaro R, Marazuela M
    PLoS ONE, 2015;10(4):e0123938.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Frozen
  3. Human T-cell leukemia virus type 2 induces survival and proliferation of CD34(+) TF-1 cells through activation of STAT1 and STAT5 by secretion of interferon-gamma and granulocyte macrophage-colony-stimulating factor.
    Authors: Bovolenta C, Pilotti E, Mauri M, Turci M, Ciancianaini P, Fisicaro P, Bertazzoni U, Poli G, Casoli C
    Blood, 2002;99(1):224-31.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
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