Flt-3 Ligand Antibody [Unconjugated]

Flt‑3 Ligand was detected in immersion fixed feline peripheral blood mononuclear cells (PBMCs) using Goat Anti-FelineFlt‑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.

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-FelineFlt-3 Ligand Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2467). The ND50 is typically 0.1-0.4 µg/mL.

• Reactivity: Fe
• Applications: WB, B/N, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Flt-3 Ligand Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant feline Flt-3 Ligand
  Ser27-Pro185
  Accession # NP_001009842
• 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: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: FLT3LG
• Purity: Immunogen affinity purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Western Blot 0.1 ug/mL
  • Immunocytochemistry 5-15 ug/mL
  • Neutralization 0.1-0.4 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Purity: Immunogen affinity purified
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Flt-3 Ligand Antibody [Unconjugated]

• FL
• Flt3 ligand
• Flt-3 Ligand
• Flt3L
• FLT3LG
• fms-related tyrosine kinase 3 ligand
• SL cytokine

Background

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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: FLT3LG
• Uniprot:
  • NP_001009842()