hh Mouse Flt-3 Ligand Antibody AF427: R&D Systems

Mouse Flt-3 Ligand Antibody

(3 citations)   
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse Flt-3 Ligand in ELISAs and Western blots. In sandwich immunoassays, less than 0.1% cross-reactivity with recombinant human Flt-3 Ligand is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant mouse Flt-3 Ligand
    Gly27-Arg188
    Accession # P49772
  • 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 Mouse Flt‑3 Ligand (Catalog # 427-FL)
  • Immunocytochemistry
    5-15 µg/mL
    See below
    • Mouse Flt-3 Ligand Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    0.2-0.8 µg/mL 
    Mouse Flt‑3 Ligand Antibody (Catalog # AF427)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Mouse Flt‑3 Ligand Biotinylated Antibody (Catalog # BAF427)
  • ELISA Standard
     
    Recombinant Mouse Flt-3 Ligand Protein (Catalog # 427-FL)
  • Neutralization
    Measured by its ability to neutralize Flt‑3 Ligand-induced proliferation in BaF3 mouse pro‑B cell line transfected with mouse Flt‑3. Hannum, C. et al. (1994) Nature 368:643. The Neutralization Dose (ND50) is typically 0.05-0.25 µg/mL in the presence of 10 ng/mL Recombinant Mouse Flt‑3 Ligand.
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 Flt‑3 Ligand and Neutralization by Mouse
Flt‑3 Ligand Antibody.
Recombinant Mouse Flt‑3 Ligand (Catalog # 427-FL) stimulates proliferation in BaF3 mouse pro‑B cell line transfected with mouse Flt‑3 in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Mouse Flt‑3 Ligand (10 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse Flt‑3 Ligand Antigen Affinity-purified Polyclonal Antibody (Catalog # AF427). The ND50 is typically 0.05‑0.25 µg/mL.
Immunocytochemistry
Flt‑3 Ligand in HT‑2 Mouse Cell Line. Flt‑3 Ligand was detected in immersion fixed HT‑2 mouse T cell line using Goat Anti-Mouse Flt‑3 Ligand Antigen Affinity-purified Polyclonal Antibody (Catalog # AF427) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
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: Flt-3 Ligand

Flt-3 Ligand, also known as FL, is an alpha -helical cytokine that promotes the differentiation of multiple hematopoietic cell lineages (1‑3). Mature mouse Flt-3 Ligand consists of a 161 amino acid (aa) extracellular domain (ECD) with a cytokine-like domain and a juxtamembrane tether region, a 21 aa transmembrane segment, and a 22 aa cytoplasmic tail (4‑6). Within the ECD, mouse Flt-3 Ligand shares 71% and 81% aa sequence identity with human and rat Flt-3 Ligand, respectively. Mouse and human Flt-3 Ligand show cross-species activity (4, 5, 7). Flt-3 Ligand is expressed as a noncovalently-linked dimer by T cells and bone marrow and thymic fibroblasts (1, 8). Each 36 kDa chain carries approximately 12 kDa of N- and O-linked carbohydrates (8). Alternate splicing and proteolytic cleavage of the transmembrane form can generate a soluble 30 kDa fragment that includes the cytokine domain (4, 8). Alternate splicing of mouse Flt-3 Ligand also generates a membrane-associated isoform with a 57 aa substitution following the cytokine domain (4, 5, 8, 9). Both transmembrane and soluble Flt-3 Ligand signal through the tyrosine kinase receptor Flt-3/Flk-2 (3‑6). Flt-3 Ligand induces the expansion of monocytes and immature dendritic cells as well as early B cell lineage differentiation (2, 10). It synergizes with IL-3, GM-CSF, and SCF to promote the mobilization and myeloid differentiation of hematopoietic stem cells (4, 5, 7). It cooperates with IL-2, -6, -7, and -15 to induce NK cell development and with IL-3, -7, and -11 to induce terminal B cell maturation (1, 11). Animal studies also show Flt-3 Ligand to reduce the severity of experimentally induced allergic inflammation (12).

  • References:
    1. Wodnar-Filipowicz, A. (2003) News Physiol. Sci. 18:247.
    2. Dong, J. et al. (2002) Cancer Biol. Ther. 1:486.
    3. Gilliland, D.G. and J.D. Griffin (2002) Blood 100:1532.
    4. Hannum, C. et al. (1994) Nature 368:643.
    5. Lyman, S.D. et al. (1993) Cell 75:1157.
    6. Savvides, S.N. et al. (2000) Nat. Struct. Biol. 7:486.
    7. Lyman, S.D. et al. (1994) Blood 83:2795.
    8. McClanahan, T. et al. (1996) Blood 88:3371.
    9. Lyman, S.D. et al. (1995) Oncogene 10:149.
    10. Diener, K.R. et al. (2008) Exp. Hematol. 36:51.
    11. Farag, S.S. and M.A. Caligiuri (2006) Blood Rev. 20:123.
    12. Edwan, J.H. et al. (2004) J. Immunol. 172:5016.
  • Long Name:
    fms-like Tyrosine Kinase 3 Ligand
  • Entrez Gene IDs:
    2323 (Human); 14256 (Mouse); 493796 (Feline)
  • Alternate Names:
    FL; Flt3 ligand; Flt-3 Ligand; Flt3L; FLT3LG; fms-related tyrosine kinase 3 ligand; SL cytokine
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. Flt3 permits survival during infection by rendering dendritic cells competent to activate NK cells.
    Authors: Eidenschenk C, Crozat K, Krebs P, Arens R, Popkin D, Arnold CN, Blasius AL, Benedict CA, Moresco EM, Xia Y, Beutler B
    Proc. Natl. Acad. Sci. U.S.A., 2010;107(21):9759-64.
    Species: Mouse
    Sample Type: In Vivo
    Application: In vivo
  2. Modulation of marrow proliferation and chemosensitivity by tumor-produced cytokines from syngeneic pancreatic tumor lines.
    Authors: Blumenthal RD, Reising A, Leon E
    Clin. Cancer Res., 2002;8(5):1301-9.
    Species: Mouse
    Sample Type: Whole Cells
    Application: Neut
  3. A secreted and LIF-mediated stromal cell-derived activity that promotes ex vivo expansion of human hematopoietic stem cells.
    Authors: Shih CC, Hu MC, Hu J
    Blood, 2000;95(6):1957-66.
    Species: Mouse
    Sample Type: Whole Cells
    Application: Neut
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