Mouse/Rat Flt-3 Ligand Quantikine ELISA Kit

(14 citations)   
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Serum (50 uL)
  • Sensitivity
    5 pg/mL
  • Assay Range
    31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum)
  • Specificity
    Natural and recombinant mouse Flt-3 Ligand and natural rat Flt-3 Ligand
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Product Summary
The Quantikine Mouse/Rat Flt-3 Ligand Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse or rat Flt-3 Ligand levels in cell culture supernates and serum. It contains NS0-expressed recombinant mouse Flt-3 Ligand and antibodies raised against the recombinant protein. Results obtained for naturally occurring mouse or rat Flt-3 Ligand showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values of natural mouse/rat Flt-3 Ligand.

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, Serum
Intra-Assay Precision Inter-Assay Precision
Standard Deviation3.910.


The recovery of mouse Flt-3 Ligand spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Mouse Cell Culture Supernates (n=5) 99 88-107
Mouse Serum (n=5) 95 82-106
Rat Cell Culture Supernates (n=5) 108 99-119
Rat Serum (n=5) 95 86-114
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of mouse Flt-3 Ligand in each matrix were diluted with Calibrator Diluent and then assayed.
Mouse/Rat Flt-3 Ligand Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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. 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. 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. Flt-3 Ligand is expressed as a noncovalently-linked dimer by T cells and bone marrow and thymic fibroblasts. Each 36 kDa chain carries approximately 12 kDa of N- and O-linked carbohydrates. Alternate splicing and proteolytic cleavage of the transmembrane form can generate a soluble 30 kDa fragment that includes the cytokine domain. 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. Both transmembrane and soluble Flt-3 Ligand signal through the tyrosine kinase receptor Flt-3/Flk-2. Flt-3 Ligand induces the expansion of monocytes and immature dendritic cells as well as early B cell lineage differentiation. It synergizes with IL-3, GM-CSF, and SCF to promote the mobilization and myeloid differentiation of hematopoietic stem cells. 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. Animal studies also show Flt-3 Ligand to reduce the severity of experimentally induced allergic inflammation.
    • Long Name
      fms-like Tyrosine Kinase 3 Ligand
    • Entrez Gene IDs
      2323 (Human); 14256 (Mouse); 493796 (Feline);
    • Alternate Names
      FL; Flt3 ligand; Flt3L; FLT3LG; fms-related tyrosine kinase 3 ligand; SL cytokine;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 50 µL Assay Diluent
    4.   Add 50 µL of Assay Diluent to each well.

    5. 50 µL Standard, Control, or Sample
    6.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
    7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

    8. 100 µL Conjugate
    9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
    10.   Aspirate and wash 5 times.

    11. 100 µL Substrate Solution
    12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

    13. 100 µL Stop Solution
    14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

    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.

    Showing Results 1 - 10 of 14
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    Sample Type
    1. Threshold levels of Flt3-ligand are required for the generation and survival of lymphoid progenitors and B-cell precursors.
      Authors: Dolence JJ, Gwin K, Frank E, Medina KL
      Eur. J. Immunol., 2012;41(2):324-34.
      Species: Mouse
      Sample Type: Serum
    2. 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;107(21):9759-64.
      Species: Mouse
      Sample Type: Serum
    3. Homeostasis of dendritic cells in lymphoid organs is controlled by regulation of their precursors via a feedback loop.
      Authors: Hochweller K, Miloud T, Striegler J, Naik S, Hammerling GJ, Garbi N
      Blood, 2009;114(20):4411-21.
      Species: Mouse
      Sample Type: Serum
    4. Ectodomain shedding of FLT3 ligand is mediated by TNF-alpha converting enzyme.
      Authors: Horiuchi K, Morioka H, Takaishi H, Akiyama H, Blobel CP, Toyama Y
      J. Immunol., 2009;182(12):7408-14.
      Species: Mouse
      Sample Type: Serum
    5. Knock-in of an internal tandem duplication mutation into murine FLT3 confers myeloproliferative disease in a mouse model.
      Authors: Li L, Piloto O, Nguyen HB, Greenberg K, Takamiya K, Racke F, Huso D, Small D
      Blood, 2008;111(7):3849-58.
      Species: Mouse
      Sample Type: Serum
    6. IL-7 induces myelopoiesis and erythropoiesis.
      Authors: Aiello FB, Keller JR, Klarmann KD, Dranoff G, Mazzucchelli R, Durum SK
      J. Immunol., 2007;178(3):1553-63.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    7. Effects of DNA- and Mycobacterium bovis BCG-based delivery of the Flt3 ligand on protective immunity to Mycobacterium tuberculosis.
      Authors: Triccas JA, Shklovskaya E, Spratt J, Ryan AA, Palendira U, Fazekas de St Groth B, Britton WJ
      Infect. Immun., 2007;75(11):5368-75.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    8. Nanosized bioceramic particles could function as efficient gene delivery vehicles with target specificity for the spleen.
      Authors: Tan K, Cheang P, Ho IA, Lam PY, Hui KM
      Gene Ther., 2007;14(10):828-35.
      Species: Mouse
      Sample Type: Serum
    9. Fms-like tyrosine kinase 3 ligand recruits plasmacytoid dendritic cells to the brain.
      Authors: Curtin JF, King GD, Barcia C, Liu C, Hubert FX, Guillonneau C, Josien R, Anegon I, Lowenstein PR, Castro MG
      J. Immunol., 2006;176(6):3566-77.
      Species: Rat
      Sample Type: Serum
    10. The long-term but not the short-term antiviral effect of IFN-alpha depends on Flt3 ligand and pDC.
      Authors: Vollstedt S, O&amp;amp;apos;Keeffe M, Ryf B, Glanzmann B, Hochrein H, Suter M
      Eur. J. Immunol., 2006;36(5):1231-40.
      Species: Mouse
      Sample Type: Serum
    11. Inflammatory and anti-glioma effects of an adenovirus expressing human soluble Fms-like tyrosine kinase 3 ligand (hsFlt3L): treatment with hsFlt3L inhibits intracranial glioma progression.
      Authors: Ali S, Curtin JF, Zirger JM, Xiong W, King GD, Barcia C, Liu C, Puntel M, Goverdhana S, Lowenstein PR, Castro MG
      Mol. Ther., 2004;10(6):1071-84.
      Species: Rat
      Sample Type: Tissue Homogenates
    12. Conditional expression of murine Flt3 ligand leads to expansion of multiple dendritic cell subsets in peripheral blood and tissues of transgenic mice.
      Authors: Manfra DJ, Chen SC, Jensen KK, Fine JS, Wiekowski MT, Lira SA
      J. Immunol., 2003;170(6):2843-52.
      Species: Mouse
      Sample Type: Serum
    13. Endogenous granulocyte-macrophage colony-stimulating factor overexpression in vivo results in the long-term recruitment of a distinct dendritic cell population with enhanced immunostimulatory function.
      Authors: Miller G, Pillarisetty VG, Shah AB, Lahrs S, Xing Z, DeMatteo RP
      J. Immunol., 2002;169(6):2875-85.
      Species: Mouse
      Sample Type: Serum
    14. 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;8(5):1301-9.
      Species: Mouse
      Sample Type: Cell Culture Supernates
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