Human IL-3 Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human IL-3 in direct ELISAs and Western blots. In direct ELISA, less than 1% cross-reactivity with recombinant mouse IL-3 and recombinant rat IL-3 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Protein A or G purified
  • Immunogen
    E. coli-derived recombinant human IL-3
    Ala20-Phe152
    Accession # Q6GS87
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
  • 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 (Catalog # 203-IL)
  • Immunocytochemistry
    5-15 µg/mL
    Immersion fixed human peripheral blood lymphocytes
  • 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 0.4-2 µg/mL in the presence of 1.25 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
Cell Proliferation Induced by IL‑3 and Neutral­ization by Human IL‑3 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 (1.25 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human IL‑3 Polyclonal Antibody (Catalog # AB-203-NA). The ND50 is typically 0.4‑2 µg/mL.
Preparation and Storage
  • Reconstitution
    Reconstitute at 1 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it 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-3

LAG-3 (Lymphocyte activation gene-3), also known as CD223, is a member of the immunoglobulin superfamily (IgSF). The mature LAG-3 protein is a 496 amino acid (aa) membrane protein with a 421 aa extracellular region which contains four IgSF domains, a 21 aa transmembrane region and a 54 aa cytoplasmic region. LAG-3 and CD4 molecules share <20% aa sequence homology but have a similar structure (1, 2). Both molecules bind to MHC class II. LAG-3 binds to MHC class II with higher affinity compared to CD4. Both LAG-3 and CD4 genes are located on the distal part of the short arm of chromosome 12.

LAG-3 is an activation-induced molecule, expressed on activated T cells and NK cells, but not on resting T cells. Studies using LAG-3 -/- mice have shown significant delay of T cell apoptosis following antigen stimulation and increased size of memory T cells pool following infection (3, 4). It also has been reported that anti-LAG-3 antibodies up-regulate T cell activation by blocking interaction of LAG-3 and MHC class II. The study has demonstrated that LAG-3 is selectively expressed on activated CD4+CD25+ TReg cells and plays a role in their suppressive activity (5). This evidence indicated, unlike the interaction of CD4 with MHC class II that plays a positive role in T cell activation, LAG-3 binds to MHC class II and negatively regulates T cell activation through LAG-3 signaling. On the other hand, studies have shown that binding of LAG-3 to MHC class II molecules on antigen presenting cells induce maturation of dendritic cells and cytokine secretion by monocytes through MHC class II signal transduction (6). Taken together, LAG-3 may have two major functions, it negatively regulates T cells activation through LAG-3 signaling and stimulates antigen presenting cells which express MHC class II.

  • References:
    1. Triebel, F. et al. (1990) J. Exp. Med. 171:1393.
    2. Baixeras, E. et al. (1992) J. Exp. Med 176:327.
    3. Workman, C.J. and D.A. Vignali (2003) Eur. J. Immunol. 33:970.
    4. Workman, C.J. et al. (2004) J. Immunol. 172:5450.
    5. Huang, C.T. et al. (2004) Immunity 21:503.
    6. Andreae, S. et al. (2003) Blood 102:2130.
  • Long Name:
    Interleukin 3
  • Entrez Gene IDs:
    3562 (Human); 16187 (Mouse); 24495 (Rat)
  • Alternate Names:
    Hematopoietic growth factor; IL3; IL-3; IL-3MGC79398; interleukin 3 (colony-stimulating factor, multiple); interleukin-3; Mast cell growth factor; mast-cell growth factor; MCGFMGC79399; MULTI-CSF; multilineage-colony-stimulating factor; Multipotential colony-stimulating factor; P-cell stimulating factor; P-cell-stimulating factor
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