Feline Flt-3 Ligand/FLT3L Antibody

Catalog #: AF2467 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF2467
AF2467-SP

Cell Proliferation Induced by Flt-3 Ligand/FLT3L and Neutralization by Feline Flt-3 Ligand/FLT3L Antibody.

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Feline Flt-3 Ligand/FLT3L Antibody Summary

Species Reactivity

Feline

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.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

Mouse myeloma cell line NS0-derived recombinant feline Flt-3 Ligand/FLT3L
Ser27-Pro185
Accession # NP_001009842

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.

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/FLT3L (Catalog # 2467-FL)

Immunocytochemistry

5-15 µg/mL

See below

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.

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.

Scientific Data

Neutralization

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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.

Immunocytochemistry

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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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile 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.

Background: Flt-3 Ligand/FLT3L

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