Feline Flt-3 Ligand Antibody

  
  • Species Reactivity
    Feline
  • Specificity
    Detects feline Flt‑3 Ligand in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 10% cross‑reactivity with recombinant human Flt-3 Ligand and recombinant mouse Flt-3 Ligand is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant feline Flt-3 Ligand
    Ser27-Pro185
    Accession # NP_001009842
  • 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 Feline Flt‑3 Ligand (Catalog # 2467-FL)
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize Flt‑3 Ligand-induced proliferation in BaFlt3 mouse pro‑B cell line transfected with mouse Flt‑3. Hannum, C. et al. (1994) Nature 368:643. The Neutralization Dose (ND50) is typically 0.1‑0.4 µg/mL in the presence of 2 ng/mL Recombinant Feline 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 Feline Flt‑3 Ligand Antibody. Recombinant Feline Flt‑3 Ligand (Catalog # 2467-FL) stimulates proliferation in BaFlt3 mouse pro‑B cell line transfected with mouse Flt‑3 in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Feline Flt‑3 Ligand (2 ng/mL) is neutralized (green line) by increasing concen­trations of Goat Anti-Feline
Flt-3 Ligand Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2467). The ND50 is typically 0.1-0.4 µg/mL.

Immunocytochemistry

Flt‑3 Ligand in Feline PBMCs. Flt‑3 Ligand was detected in immersion fixed feline peripheral blood mononuclear cells (PBMCs) using Goat Anti-Feline
Flt‑3 Ligand Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2467) at 15 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 feline Flt-3 Ligand consists of a 156 amino acid (aa) extracellular region with a cytokine-like domain and a juxtamembrane tether region, a 21 aa transmembrane segment, and an 88 aa cytoplasmic tail (4, 5). Within the ECD, feline Flt-3 Ligand shares 92%, 81%, 70%, and 63% aa sequence identity with canine, human, mouse, and rat Flt-3 Ligand, respectively. Flt-3 Ligand is expressed as a noncovalently-linked dimer by T cells and bone marrow and thymic fibroblasts (1, 6). Each 36 kDa chain carries approximately 12 kDa of N- and O-linked carbohydrates (6). Alternate splicing and proteolytic cleavage of the transmembrane form can generate a soluble 30 kDa fragment that includes the cytokine domain (6, 7). Multiple splice forms of canine, human, and mouse Flt-3 Ligand have been described and may also exist in feline (4, 6‑10). Both transmembrane and soluble Flt-3 Ligand signal through the tyrosine kinase receptor Flt-3/Flk-2 (3, 5, 7, 10). Flt-3 Ligand induces the expansion of monocytes and immature dendritic cells as well as early B cell lineage differentiation (2, 11). It synergizes with IL-3, GM-CSF, and SCF to promote the mobilization and myeloid differentiation of hematopoietic stem cells (7, 8, 10). 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, 12). Animal studies also show Flt-3 Ligand to reduce the severity of experimentally induced allergic inflammation (13).

  • 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. Yang, S. and G.-K. Sim (2000) DNA Seq. 11:163.
    5. Savvides, S.N. et al. (2000) Nat. Struct. Biol. 7:486.
    6. McClanahan, T. et al. (1996) Blood 88:3371.
    7. Hannum, C. et al. (1994) Nature 368:643.
    8. Lyman, S.D. et al. (1994) Blood 83:2795.
    9. Lyman, S.D. et al. (1995) Oncogene 10:149.
    10. Lyman, S.D. et al. (1993) Cell 75:1157.
    11. Diener, K.R. et al. (2008) Exp. Hematol. 36:51.
    12. Farag, S.S. and M.A. Caligiuri (2006) Blood Rev. 20:123.
    13. 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
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