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Mouse Flt-3 Ligand/FLT3L Antibody

Catalog #: AF427
8 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF427
AF427-SP
Cell Proliferation Induced by Flt-3 Ligand/FLT3L and Neutralization by Mouse Flt-3 Ligand/FLT3L Antibody.
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Mouse Flt-3 Ligand/FLT3L Antibody Summary

Species Reactivity
Mouse
Specificity
Detects mouse Flt-3 Ligand/FLT3L in ELISAs and Western blots. In sandwich immunoassays, less than 0.1% cross-reactivity with recombinant human Flt-3 Ligand/FLT3L is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant mouse Flt-3 Ligand/FLT3L
Gly27-Arg188
Accession # P49772
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 Mouse Flt-3 Ligand/FLT3L (Catalog # 427-FL)
Immunocytochemistry
5-15 µg/mL
See below

Mouse Flt-3 Ligand/FLT3L Sandwich Immunoassay

Recommended Concentration
Reagent
ELISA Capture (Matched Antibody Pair)
0.2-0.8 µg/mL 

Use in combination with:

Detection Reagent: Mouse Flt-3 Ligand/FLT3L Biotinylated Antibody (Catalog # BAF427)

Standard: Recombinant Mouse Flt-3 Ligand/FLT3L Protein (Catalog # 427-FL)

Neutralization
Measured by its ability to neutralize Flt‑3 Ligand/FLT3L-induced proliferation in BaF3 mouse pro‑B cell line transfected with mouse Flt‑3. Hannum, C. et al. (1994) Nature 368:643. The Neutralization Dose (ND50) is typically 0.05-0.25 µg/mL in the presence of 10 ng/mL Recombinant Mouse 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 Cell Proliferation Induced by Flt-3 Ligand/FLT3L and Neutralization by Mouse Flt-3 Ligand/FLT3L Antibody. View Larger

Cell Proliferation Induced by Flt-3 Ligand/FLT3L and Neutralization by Mouse Flt-3 Ligand/FLT3L Antibody. Recombinant Mouse Flt-3 Ligand/FLT3L (Catalog # 427-FL) stimulates proliferation in BaF3 mouse pro-B cell line transfected with mouse Flt-3 in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Mouse Flt-3 Ligand/FLT3L (10 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse Flt-3 Ligand/FLT3L Antigen Affinity-purified Polyclonal Antibody (Catalog # AF427). The ND50 is typically 0.05-0.25 µg/mL.

Immunocytochemistry Flt-3 Ligand/FLT3L antibody in HT-2 Mouse Cell Line by Immunocytochemistry (ICC). View Larger

Flt-3 Ligand/FLT3L in HT‑2 Mouse Cell Line. Flt-3 Ligand/FLT3L was detected in immersion fixed HT-2 mouse T cell line using Goat Anti-Mouse Flt-3 Ligand/FLT3L Antigen Affinity-purified Polyclonal Antibody (Catalog # AF427) at 5 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.

Flow Cytometry Detection of Mouse Flt-3 Ligand/FLT3L by Flow Cytometry View Larger

Detection of Mouse Flt-3 Ligand/FLT3L by Flow Cytometry Flt3L-producing tumors do not expand mature ILCs in periphery. Mice were injected with 2 × 106 B16-Flt3L or B16/PBS as a control. Two weeks after tumor injection, ILCs were analyzed by FACS in the lungs, small intestine and colonic lamina propria. (a) Representative dot plots showing NK1.1+T-bet+Eomes− ILC1, NK1.1+T-bet+Eomes+ cNK, GATA-3+ ILC2, ROR gamma t+ ILC3 and NKp46+ and CCR6+ ILC3 subsets gated on CD45+ Lin (CD3, CD19)neg CD90+ cells in the colonic lamina propria and (b) quantification of absolute numbers (n = 5 to 7/group, 3 experiments). Representative dot plots (c) and absolute numbers (d) of ILC2 and ILC3 in the small intestine lamina propria and lungs of B16-Flt3L and B16/PBS injected mice (n = 5 to 8/group, 3 experiments). (e) Representative dot plots of IL22+ ROR gamma t+ ILC3 and IL-5+ GATA-3+ ILC2 in the small intestine of B16-Flt3L and B16 injected mice (n = 3 to 4, 2 experiments). *P < 0.05; ns, not significant; Student’s t-test (b,d,e). Error bars represent SEM in all panels. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29317685), licensed under a CC-BY license. Not internally tested by R&D Systems.

Flow Cytometry Detection of Mouse Flt-3 Ligand/FLT3L by Flow Cytometry View Larger

Detection of Mouse Flt-3 Ligand/FLT3L by Flow Cytometry Flt3L-producing tumors expand CHILPs in the BM. Mice were injected with 2 × 106 B16-Flt3L or B16/PBS as a control. Two weeks after tumor injection, Flt3L serum levels were analyzed by ELISA and CHILPs and ILC2P were analyzed by FACS on BM single-cell suspension. (a) Flt3L serum levels in B16/PBS and B16-Flt3L injected mice (n = 3/group, 2 experiments). Representative dot plots (b) and total number (c) of Lin− alpha 4 beta 7+ cells in the BM of wild-type mice injected with B16-Flt3L or B16/PBS (n = 6/group, 3 experiments). Representative dot plots (d) and absolute numbers (e) of CHILPs and ILC2P in the BM of Id2Gfp/+ mice injected with B16-Flt3L or B16/PBS (n = 6/group, 3 experiments). (f) Correlation plot between Flt3L serum levels determined by ELISA and BM CHILPs absolute numbers determined by FACS in mice treated with B16-Flt3L or B16 (n = 6, 2 experiments). Representative dot plots (g) and absolute numbers (h) of ILCP in the bone marrow of wild type mice injected with B16-Flt3L or B16 cells (n = 5 to 7/group, 3 experiments). *P < 0.05; **P < 0.01; ns, not significant; Student’s t-test (c,e,h), Linear regression with Pearson’s correlation analysis (f). Error bars represent SEM in all panels. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29317685), licensed under a CC-BY license. Not internally tested by R&D Systems.

Flow Cytometry Detection of Mouse Flt-3 Ligand/FLT3L by Flow Cytometry View Larger

Detection of Mouse Flt-3 Ligand/FLT3L by Flow Cytometry Flt3L-producing tumors do not expand mature ILCs in periphery. Mice were injected with 2 × 106 B16-Flt3L or B16/PBS as a control. Two weeks after tumor injection, ILCs were analyzed by FACS in the lungs, small intestine and colonic lamina propria. (a) Representative dot plots showing NK1.1+T-bet+Eomes− ILC1, NK1.1+T-bet+Eomes+ cNK, GATA-3+ ILC2, ROR gamma t+ ILC3 and NKp46+ and CCR6+ ILC3 subsets gated on CD45+ Lin (CD3, CD19)neg CD90+ cells in the colonic lamina propria and (b) quantification of absolute numbers (n = 5 to 7/group, 3 experiments). Representative dot plots (c) and absolute numbers (d) of ILC2 and ILC3 in the small intestine lamina propria and lungs of B16-Flt3L and B16/PBS injected mice (n = 5 to 8/group, 3 experiments). (e) Representative dot plots of IL22+ ROR gamma t+ ILC3 and IL-5+ GATA-3+ ILC2 in the small intestine of B16-Flt3L and B16 injected mice (n = 3 to 4, 2 experiments). *P < 0.05; ns, not significant; Student’s t-test (b,d,e). Error bars represent SEM in all panels. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29317685), licensed under a CC-BY license. Not internally tested by R&D Systems.

ELISA Detection of Mouse Flt-3 Ligand/FLT3L by ELISA View Larger

Detection of Mouse Flt-3 Ligand/FLT3L by ELISA Flt3L-producing tumors expand CHILPs in the BM. Mice were injected with 2 × 106 B16-Flt3L or B16/PBS as a control. Two weeks after tumor injection, Flt3L serum levels were analyzed by ELISA and CHILPs and ILC2P were analyzed by FACS on BM single-cell suspension. (a) Flt3L serum levels in B16/PBS and B16-Flt3L injected mice (n = 3/group, 2 experiments). Representative dot plots (b) and total number (c) of Lin− alpha 4 beta 7+ cells in the BM of wild-type mice injected with B16-Flt3L or B16/PBS (n = 6/group, 3 experiments). Representative dot plots (d) and absolute numbers (e) of CHILPs and ILC2P in the BM of Id2Gfp/+ mice injected with B16-Flt3L or B16/PBS (n = 6/group, 3 experiments). (f) Correlation plot between Flt3L serum levels determined by ELISA and BM CHILPs absolute numbers determined by FACS in mice treated with B16-Flt3L or B16 (n = 6, 2 experiments). Representative dot plots (g) and absolute numbers (h) of ILCP in the bone marrow of wild type mice injected with B16-Flt3L or B16 cells (n = 5 to 7/group, 3 experiments). *P < 0.05; **P < 0.01; ns, not significant; Student’s t-test (c,e,h), Linear regression with Pearson’s correlation analysis (f). Error bars represent SEM in all panels. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29317685), licensed under a CC-BY license. Not internally tested by R&D Systems.

Flow Cytometry Detection of Mouse Flt-3 Ligand/FLT3L by Flow Cytometry View Larger

Detection of Mouse Flt-3 Ligand/FLT3L by Flow Cytometry Flt3L-producing tumors do not expand mature ILCs in periphery. Mice were injected with 2 × 106 B16-Flt3L or B16/PBS as a control. Two weeks after tumor injection, ILCs were analyzed by FACS in the lungs, small intestine and colonic lamina propria. (a) Representative dot plots showing NK1.1+T-bet+Eomes− ILC1, NK1.1+T-bet+Eomes+ cNK, GATA-3+ ILC2, ROR gamma t+ ILC3 and NKp46+ and CCR6+ ILC3 subsets gated on CD45+ Lin (CD3, CD19)neg CD90+ cells in the colonic lamina propria and (b) quantification of absolute numbers (n = 5 to 7/group, 3 experiments). Representative dot plots (c) and absolute numbers (d) of ILC2 and ILC3 in the small intestine lamina propria and lungs of B16-Flt3L and B16/PBS injected mice (n = 5 to 8/group, 3 experiments). (e) Representative dot plots of IL22+ ROR gamma t+ ILC3 and IL-5+ GATA-3+ ILC2 in the small intestine of B16-Flt3L and B16 injected mice (n = 3 to 4, 2 experiments). *P < 0.05; ns, not significant; Student’s t-test (b,d,e). Error bars represent SEM in all panels. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29317685), licensed under a CC-BY license. Not internally tested by R&D Systems.

Flow Cytometry Detection of Mouse Flt-3 Ligand/FLT3L by Flow Cytometry View Larger

Detection of Mouse Flt-3 Ligand/FLT3L by Flow Cytometry Flt3L-producing tumors expand CHILPs in the BM. Mice were injected with 2 × 106 B16-Flt3L or B16/PBS as a control. Two weeks after tumor injection, Flt3L serum levels were analyzed by ELISA and CHILPs and ILC2P were analyzed by FACS on BM single-cell suspension. (a) Flt3L serum levels in B16/PBS and B16-Flt3L injected mice (n = 3/group, 2 experiments). Representative dot plots (b) and total number (c) of Lin− alpha 4 beta 7+ cells in the BM of wild-type mice injected with B16-Flt3L or B16/PBS (n = 6/group, 3 experiments). Representative dot plots (d) and absolute numbers (e) of CHILPs and ILC2P in the BM of Id2Gfp/+ mice injected with B16-Flt3L or B16/PBS (n = 6/group, 3 experiments). (f) Correlation plot between Flt3L serum levels determined by ELISA and BM CHILPs absolute numbers determined by FACS in mice treated with B16-Flt3L or B16 (n = 6, 2 experiments). Representative dot plots (g) and absolute numbers (h) of ILCP in the bone marrow of wild type mice injected with B16-Flt3L or B16 cells (n = 5 to 7/group, 3 experiments). *P < 0.05; **P < 0.01; ns, not significant; Student’s t-test (c,e,h), Linear regression with Pearson’s correlation analysis (f). Error bars represent SEM in all panels. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29317685), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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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 mouse Flt-3 Ligand consists of a 161 amino acid (aa) extracellular domain (ECD) with a cytokine-like domain and a juxtamembrane tether region, a 21 aa transmembrane segment, and a 22 aa cytoplasmic tail (4‑6). Within the ECD, mouse Flt-3 Ligand shares 71% and 81% aa sequence identity with human and rat Flt-3 Ligand, respectively. Mouse and human Flt-3 Ligand show cross-species activity (4, 5, 7). 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 mouse Flt-3 Ligand also generates a membrane-associated isoform with a 57 aa substitution following the cytokine domain (4, 5, 8, 9). Both transmembrane and soluble Flt-3 Ligand signal through the tyrosine kinase receptor Flt-3/Flk-2 (3‑6). Flt-3 Ligand induces the expansion of monocytes and immature dendritic cells as well as early B cell lineage differentiation (2, 10). It synergizes with IL-3, GM-CSF, and SCF to promote the mobilization and myeloid differentiation of hematopoietic stem cells (4, 5, 7). 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, 11). Animal studies also show Flt-3 Ligand to reduce the severity of experimentally induced allergic inflammation (12).

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. (1993) Cell 75:1157.
  6. Savvides, S.N. et al. (2000) Nat. Struct. Biol. 7:486.
  7. Lyman, S.D. et al. (1994) Blood 83:2795.
  8. McClanahan, T. et al. (1996) Blood 88:3371.
  9. Lyman, S.D. et al. (1995) Oncogene 10:149.
  10. Diener, K.R. et al. (2008) Exp. Hematol. 36:51.
  11. Farag, S.S. and M.A. Caligiuri (2006) Blood Rev. 20:123.
  12. 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); 102125470 (Cynomolgus Monkey); 493796 (Feline)
Alternate Names
FL; FLG3L; Flt3 ligand; Flt-3 Ligand; Flt3L; FLT3LG; fms-related tyrosine kinase 3 ligand; SL cytokine

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Citations for Mouse Flt-3 Ligand/FLT3L Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

8 Citations: Showing 1 - 8
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  1. A natural killer–dendritic cell axis defines checkpoint therapy–responsive tumor microenvironments
    Authors: Kevin C. Barry, Joy Hsu, Miranda L. Broz, Francisco J. Cueto, Mikhail Binnewies, Alexis J. Combes et al.
    Nature Medicine
  2. Route of Vaccine Administration Influences the Impact of Fms-Like Tyrosine Kinase 3 Ligand (Flt3L) on Chlamydial-Specific Protective Immune Responses
    Authors: Roshan Pais, Yusuf Omosun, Joseph U. Igietseme, Kohtaro Fujihashi, Francis O. Eko
    Frontiers in Immunology
  3. Hto, Tritiated Amino Acid Exposure and External Exposure Induce Differential Effects on Hematopoiesis and Iron Metabolism
    Authors: JM Bertho, D Kereselidz, L Manens, C Culeux, V Magneron, J Surette, M Blimkie, L Bertrand, H Wyatt, M Souidi, I Dublineau, N Priest, JR Jourdain
    Sci Rep, 2019-12-27;9(1):19919.
    Species: Mouse
    Sample Types: Plasma
    Applications: Multiplex Assay
  4. Acutely malnourished weanling mice administered Flt3 ligand can support a cell-mediated inflammatory response
    Authors: LM Hillyer, B Woodward
    Cytokine, 2018-07-07;113(0):39-49.
    Species: Human
    Sample Types: Cell Culture Supernates
    Applications: ELISA Development (Capture), Western Blot
  5. Flt3 permits survival during infection by rendering dendritic cells competent to activate NK cells.
    Authors: Eidenschenk C, Crozat K, Krebs P, Arens R, Popkin D, Arnold CN, Blasius AL, Benedict CA, Moresco EM, Xia Y, Beutler B
    Proc. Natl. Acad. Sci. U.S.A., 2010-05-10;107(21):9759-64.
    Species: Mouse
    Sample Types: In Vivo
    Applications: Neutralization
  6. Modulation of marrow proliferation and chemosensitivity by tumor-produced cytokines from syngeneic pancreatic tumor lines.
    Authors: Blumenthal RD, Reising A, Leon E
    Clin. Cancer Res., 2002-05-01;8(5):1301-9.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Neutralization
  7. A secreted and LIF-mediated stromal cell-derived activity that promotes ex vivo expansion of human hematopoietic stem cells.
    Authors: Shih CC, Hu MC, Hu J
    Blood, 2000-03-15;95(6):1957-66.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Neutralization
  8. The Magnitude and IgG Subclass of Antibodies Elicited by Targeted DNA Vaccines Are Influenced by Specificity for APC Surface Molecules
    Authors: Ranveig Braathen, Heidi C. L. Spång, Mona M. Lindeberg, Even Fossum, Gunnveig Grødeland, Agnete B. Fredriksen et al.
    ImmunoHorizons

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