Rat IL-3 Antibody

  
  • Species Reactivity
    Rat
  • Specificity
    Detects rat IL-3 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 5% cross-reactivity with recombinant mouse IL-3 and less than 1% cross-reactivity with recombinant human IL-3 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant rat IL‑3
    Ile27-Cys169
    Accession # P97688
  • 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
    0.1 µg/mL
    Recombinant Rat IL‑3 (Catalog # 2524-RL)
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize IL‑3-induced proliferation in the M‑NFS‑60 mouse myelogenous leukemia lymphoblast cell line. Holmes, K.L. et al. (1985) Proc. Natl. Acad. Sci. USA 82:6687. The Neutralization Dose (ND50) is typically 0.75-3 µg/mL in the presence of 0.5 ng/mL Recombinant Rat 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 in Rat Splenocytes. IL‑3 was detected in immersion fixed rat splenocytes using Goat Anti-Rat IL‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2524) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (yellow; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Cell Proliferation Induced by IL‑3 and Neutralization by Rat IL‑3 Antibody. Recombinant Rat IL‑3 (Catalog # 2524-RL) stimulates proliferation in the M‑NFS‑60 mouse myelogenous leukemia lymphoblast cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Rat IL‑3 (0.5 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Rat IL‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2524). The ND50 is typically 0.75-3 µg/mL.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 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

Rat interleukin-3 (IL-3; also multi-CSF) is a 26 kDa, variably glycosylated monomeric polypeptide that belongs to the alpha -helix family of hematopoietic cytokines (1, 2). IL-3 has pleiotrophic activies on a number of hematopoietic-related cells (1, 3). The rat molecule has two alternate splice forms. The first is termed IL-3 beta and is synthesized as a 169 amino acid (aa) precursor that contains a 27 aa signal sequence and a 142 aa mature segment (1, 2). The second is called IL-3 alpha, and is identical to IL-3 beta, save for a three amino acid (Tyr-Pro-Gln) deletion at positions 56-58 (1). The beta form is considered the most common form. Each form has an alpha -helical structure with two intrachain disulfide bonds and two potential N-linked glycosylation sites. Rat IL-3 is generally considered to be species-specific in its activity. In the mature region, rat IL-3 shares 55%, 30%, and 24% aa sequence identity with mouse, human, and bovine IL-3, respectively. Cells known to express IL-3 include connective tissue mast cells, astrocytes, microglia, megakaryocytes, eosinophils, T cells, keratinocytes and thymic epithelium.

IL-3 exerts its biological activities by binding to a 70 kDa, low affinity, ligand-binding IL-3 R alpha subunit, (6) which then recruits a 120 kDa, common beta -chain, signal transducing subunit (7) to form a functional IL-3 receptor (1, 6, 7). Receptors for IL-3 are present on bone marrow progenitors, macrophages, mast cells, eosinophils, megakaryocytes, basophils, and various myeloid leukemic cells. IL-3 can stimulate the proliferation and differentiation of pluripotent hematopoietic stem cells as well as various lineage committed progenitors including those for neutrophils, macrophages, magakaryocytes, and erythroid cells. IL-3 can stimulate the growth of early B cells and mature macrophages, mast cells, eosinophils, basophils, and megakaryocytes. IL-3 augments the function activity of basophils, mast cells, eosinophils, and macrophages (1, 8). In combination with other molecules such as CD40L and or IL-4, IL-3 can stimulate production of dendritic cells (1, 2, 9, 10).

  • References:
    1. Martinez-Moczygemba, M. and D.P. Huston (2003) J. Allergy Clin. Immunol. 112:653.
    2. Mangi, M.H. and A.C. Newland (1999) Cytokines Cell. Mol. Ther. 5:87.
    3. Esandi, M. del C. et al. (1998) Gene 211:151.
    4. Cohen, D.R. et al. (1986) Nucleic Acids Res. 14:3641.
    5. Gebicke-Haerter, P.J. et al. (1994) J. Neuroimmunol. 50:203.
    6. Chritton, S.C. and M.H. Sheng (1997) GenBank Accession # NP_640353.
    7. Appel, K. et al. (1995) J. Neurosci. 15:5800.
    8. Schrader, J.W. (2001) in Cytokine Reference, Oppenhiem, J.J. and M. Feldmann (eds): Academic Press, p. 855. 
    9. Ebner, S. et al. (2002) J. Immunol. 168:6199.
    10. Buelens C. et. al. (2002) Blood 99:993.
  • 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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