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).
Key Product Details
Species Reactivity
Feline
Applications
Western Blot, Neutralization, Immunocytochemistry
Label
Unconjugated
Antibody Source
Polyclonal Goat IgG
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Product Specifications
Immunogen
Mouse myeloma cell line NS0-derived recombinant feline Flt-3 Ligand/FLT3L
Ser27-Pro185
Accession # NP_001009842
Ser27-Pro185
Accession # NP_001009842
Specificity
Detects feline Flt‑3 Ligand/FLT3L in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 10% cross‑reactivity with recombinant human Flt-3 Ligand/FLT3L and recombinant mouse Flt-3 Ligand/FLT3L is observed.
Clonality
Polyclonal
Host
Goat
Isotype
IgG
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Scientific Data Images for Feline Flt-3 Ligand/FLT3L Antibody
Cell Proliferation Induced by Flt-3 Ligand/FLT3L and Neutralization by Feline Flt-3 Ligand/FLT3L Antibody.
Recombinant Feline Flt-3 Ligand/FLT3L (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/FLT3L (2 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Feline Flt-3 Ligand/FLT3L Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2467). The ND50 is typically 0.1-0.4 µg/mL.Flt-3 Ligand/FLT3L in Feline PBMCs.
Flt-3 Ligand/FLT3L was detected in immersion fixed feline peripheral blood mononuclear cells (PBMCs) using Goat Anti-Feline Flt-3 Ligand/FLT3L 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.Applications for Feline Flt-3 Ligand/FLT3L Antibody
Application
Recommended Usage
Immunocytochemistry
5-15 µg/mL
Sample: Immersion fixed feline peripheral blood mononuclear cells (PBMCs)
Sample: Immersion fixed feline peripheral blood mononuclear cells (PBMCs)
Western Blot
0.1 µg/mL
Sample: Recombinant Feline Flt‑3 Ligand/FLT3L (Catalog # 2467-FL)
Sample: Recombinant Feline Flt‑3 Ligand/FLT3L (Catalog # 2467-FL)
Neutralization
Measured by its ability to neutralize Flt‑3 Ligand/FLT3L-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/FLT3L.
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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.
Calculators
Background: Flt-3 Ligand/FLT3L
References
- Wodnar-Filipowicz, A. (2003) News Physiol. Sci. 18:247.
- Dong, J. et al. (2002) Cancer Biol. Ther. 1:486.
- Gilliland, D.G. and J.D. Griffin (2002) Blood 100:1532.
- Yang, S. and G.-K. Sim (2000) DNA Seq. 11:163.
- Savvides, S.N. et al. (2000) Nat. Struct. Biol. 7:486.
- McClanahan, T. et al. (1996) Blood 88:3371.
- Hannum, C. et al. (1994) Nature 368:643.
- Lyman, S.D. et al. (1994) Blood 83:2795.
- Lyman, S.D. et al. (1995) Oncogene 10:149.
- Lyman, S.D. et al. (1993) Cell 75:1157.
- Diener, K.R. et al. (2008) Exp. Hematol. 36:51.
- Farag, S.S. and M.A. Caligiuri (2006) Blood Rev. 20:123.
- Edwan, J.H. et al. (2004) J. Immunol. 172:5016.
Long Name
fms-like Tyrosine Kinase 3 Ligand
Alternate Names
FLG3L, Flt3L, FLT3LG
Gene Symbol
FLT3LG
UniProt
Additional Flt-3 Ligand/FLT3L Products
Product Documents for Feline Flt-3 Ligand/FLT3L Antibody
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Feline Flt-3 Ligand/FLT3L Antibody
For research use only
Related Research Areas
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars