Recombinant Human Flt-3 Ligand GMP Protein, CF

Animal Component Free Process.
  
  • Purity
    >97%, by SDS-PAGE with silver staining, under reducing conditions.
  • Endotoxin Level
    <0.01 EU per 1 μg of the protein by the LAL method.
  • Activity
    Measured in a cell proliferation assay using BaF3 mouse pro‑B cells transfected with mouse Flt-3. The ED50 for this effect is 0.2-1 ng/mL.
    The specific activity of recombinant human Flt-3 Ligand is approximately 1.2 x 103 U/μg, which is calibrated against recombinant human Flt-3 Ligand WHO Standard (NIBSC code: 96/532).
  • Source
    Spodoptera frugiperda, Sf 9 (baculovirus)-derived Thr27-Pro185 Produced in an animal component free process (ACFP).
    Manufactured and tested under cGMP guidelines.
  • Accession #
  • N-terminal Sequence
    Analysis

    Thr-Gln-Asp-(Cys)-Ser-Phe-Gln-His-Ser-Pro

  • Predicted Molecular Mass
    17.5 kDa
  • SDS-PAGE
    17-30 kDa, reducing conditions
308-GMP
 
Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution Reconstitute at 100-200 μg/mL in 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.
  • A minimum of 6 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, ≤ -20 °C under sterile conditions after reconstitution.
Data Images
GMP-grade Recombinant Human Flt-3 Ligand (Catalog # 308-GMP) stimulates cell proliferation in the BaF3 mouse pro-B cell line transfected with mouse Flt-3. The ED50 for this effect is 0.2-1 ng/mL.
1 μg/lane of Recombinant Human GMP-grade Flt-3 Ligand (Catalog # 308-GMP) was resolved with SDS-PAGE under reducing (R) conditions and visualized by silver staining, showing major bands at 17‑24 kDa.
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 human Flt‑3 Ligand consists of a 158 amino acid (aa) extracellular domain (ECD) with a cytokine-like domain and a juxtamembrane tether region, a 21 aa transmembrane segment, and a 30 aa cytoplasmic tail (4-7). Within the ECD, human Flt‑3 Ligand shares 71% and 65% aa sequence identity with mouse and rat Flt‑3 Ligand, respectively. Human and mouse Flt‑3 Ligand show cross-species activity (4-6). 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 human Flt‑3 Ligand also generates membrane-associated isoforms that contain either a truncated cytoplasmic tail or an 85 aa substitution following the cytokine domain (4, 5, 8). Both transmembrane and soluble Flt‑3 Ligand signal through the tyrosine kinase receptor Flt-3/Flk-2 (3, 4, 6, 7). Flt‑3 Ligand induces the expansion of monocytes and immature dendritic cells as well as early B cell lineage differentiation (2, 9). It synergizes with IL-3, GM-CSF, and SCF to promote the mobilization and myeloid differentiation of hematopoietic stem cells (4-6). 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, 10). Animal studies also show
Flt‑3 Ligand to reduce the severity of experimentally induced allergic inflammation (11).

  • 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. (1994) Blood 83:2795.
    6. Lyman, S.D. et al. (1993) Cell 75:1157.
    7. Savvides, S.N. et al. (2000) Nat. Struct. Biol. 7:486.
    8. McClanahan, T. et al. (1996) Blood 88:3371.
    9. Diener, K.R. et al. (2008) Exp. Hematol. 36:51.
    10. Farag, S.S. and M.A. Caligiuri (2006) Blood Rev. 20:123.
    11. 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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